Methods for inhibiting fascin

ABSTRACT

Provided are compositions and methods for treating a condition or disorder mediated by fascin activity in a subject in need thereof which method comprises administering to the subject a therapeutically effective amount of at least one compound of any one of Formula I-a to I-n, II, II-a, II-b or III or a pharmaceutically acceptable salt thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.13/972,649, filed on Aug. 21, 2013, which claims the benefit of U.S.Provisional Patent Application No. 61/692,177, filed on Aug. 22, 2012,and U.S. Provisional Patent Application No. 61/778,015, filed on Mar.12, 2013. The entire contents of the foregoing applications are herebyincorporated herein by reference in their entireties.

GOVERNMENT FUNDING

The technology described herein was developed with funds from NationalInstitutes of Health Grant No. R01 CA136837. The United StatesGovernment has certain rights to the technology.

FIELD

The present technology relates generally to methods for treating orpreventing cancer.

BACKGROUND

In recent years, progress has been made in the treatment of cancer,particularly with the development of targeted therapeutics. However,there is very little advancement in the treatment of tumor metastasis,which remains the major cause of mortality of cancer patients. Tumormetastasis being responsible for ˜90% of all cancer deaths (1, 2).Metastasis is a multi-step process wherein a primary tumor spreads fromits initial site to secondary tissues and organs (3-5). This metastaticprocess is selective for cells that succeed in cell migration, invasion,embolization, survival in the circulation, arrest in a distant capillarybed, and extravasation into and multiplication within the organparenchyma. Failure at any of these steps could block the entiremetastatic process. Since tumor spreading is responsible for themajority of deaths of cancer patients, there is a demand for thedevelopment of therapeutic agents that inhibit tumor metastasis.

Most current treatments for metastatic cancers are aimed to kill or stopthe growth of primary cancer cells (6-8). Although tumor cell migrationand invasion are critical steps in the process of tumor metastasis(9-12), inhibitors of tumor cell migration are not presently availableto treat metastatic cancer. Therefore, it is desirable to develop smallmolecule inhibitors targeting tumor cell migration.

SUMMARY

The present technology provides a method of treating a condition ordisorder mediated by fascin activity in a subject in need thereof whichmethod comprises administering to the subject a therapeuticallyeffective amount of at least one compound of Formula I-a, I-b, II orIII, or tautomer thereof, and/or a pharmaceutically acceptable saltthereof as described herein.

In one aspect, the present technology provides a method of treating acondition or disorder mediated by fascin activity in a subject in needthereof which method comprises administering to the subject atherapeutically effective amount of at least one compound, or acomposition comprising an effective amount of at least one compound, ofFormula I-a or I-b

or a tautomer thereof, and/or a pharmaceutically acceptable saltthereof, wherein

-   -   Q¹ and Q² are independently phenyl, 5-membered heteroaryl or        6-membered heteroaryl and are fused together in Formula I-a;    -   Q³ is 6-membered unsaturated ring wherein (1) the bond between        Y¹ and Y² is a double bond, and the bond between Y³ and Y² is a        single bond, or (2) the bond between    -   Y¹ and Y² is a single bond, and the bond between Y³ and Y² is a        double bond, and wherein Q³ is fused with Q² in Formula I-b;    -   s is 0 or 1; t is 1 or 2;    -   Y¹, Y³ and Y⁵ are independently C or N; Y², Y⁴ and Y⁶ are        independently CH, CR³ or N; provided that no more than four of        Y¹, Y², Y³, Y⁴, Y⁵ and Y⁶ are N;    -   R¹ is phenyl, 5-membered heteroaryl or 6-membered heteroaryl,        wherein the phenyl, 5-membered heteroaryl or 6-membered        heteroaryl is optionally substituted with 1 to 3 R⁶;    -   one of R and R⁴ is absent or is hydrogen, halo or lower alkyl        (preferably methyl or ethyl), and the other of R and R⁴ is L²-R⁵        or L³-R⁵; or R is absent and R⁴ is —(CH₂)_(j)—R¹¹; j is 1, 2 or        3; R¹¹ is selected from the group consisting of —OH, —OR⁷, —SH,        —SR⁷, —NR¹⁰R¹⁰, cyano, nitro, —COH, —COR⁷, —CO₂H, —CO₂R⁷,        —CONR¹⁰R¹⁰, —OCOR⁷, —OCO₂R⁷, —OCONR¹⁰R¹⁰, —NR¹⁰COR¹⁰,        —NR¹⁰CO₂R¹⁰, —SOR⁷, —SO₂R⁷, —SO₂NR¹⁰R¹⁰, and —NR¹⁰SO₂R⁷;    -   X¹ is selected from the group consisting of OR⁸, NHR⁸, and SR⁸;    -   X² is selected from the group consisting of O, NR⁸, and S;    -   L¹ is selected from the group consisting —(C(R⁸)₂)_(j)—,        —(C(R⁸)₂)_(q)—C(O)—(C(R⁸)₂)_(r)—,        —(C(R⁸)₂)_(q)—C(O)N(R⁸)—(C(R⁸)₂)_(r)—,        —(C(R⁸)₂)_(q)—N(R⁸)C(O)—(C(R⁸)₂)_(r)—,        —(C(R⁸)₂)_(q)—N(R⁸)S(O)₂—(C(R⁸)₂)_(r)—,        —(CH₂)_(q)—S(O)₂N(R⁸)—(CH₂)_(r)—, —S—, —O— and —NR⁸—;    -   q is 0 or 1;    -   r is 0 or 1;    -   L² is selected from the group consisting a covalent bond,        —C(O)N(R⁸)—, —N(R⁸)C(O)—, —N(R⁸)S(O)₂—, and —S(O)₂N(R⁸)—; L³ is        ═NC(O)—, or ═NS(O)₂—;    -   each R³ is independently selected from the group consisting of        lower alkyl (preferably methyl or ethyl) and halo;    -   R⁵ is phenyl, 5-membered heteroaryl, 6-membered heteroaryl,        5-membered heterocycloalkyl or 6-membered heterocycloalkyl;        wherein the phenyl, 5-membered heteroaryl or 6-membered        heteroaryl is optionally substituted with 1 to 4 R², wherein        each R² is independently selected from the group consisting of        lower alkyl, lower haloalkyl, —OH, —OR⁷, —SH, —SR⁷, —NR¹⁰R¹⁰,        halo, cyano, nitro, —COH, —COR⁷, —CO₂H, —CO₂R⁷, —CONR¹OR¹⁰,        —OCOR⁷, —OCO₂R⁷, —OCONR¹⁰R¹⁰, —NR¹⁰COR¹⁰, —NR¹⁰CO₂R¹⁰, —SOR⁷,        —SO₂R⁷, —SO₂NR¹⁰R¹⁰, and —NR¹⁰SO₂R⁷;    -   each R⁶ is independently selected from the group consisting of        halo and lower alkyl (preferably methyl or ethyl) optionally        substituted with 1-3 halo; or two adjacent R⁶ on a phenyl ring        form a 5- or 6-membered cycloalkyl or heterocycloalkyl fused        with the phenyl ring;    -   R⁷ is lower alkyl (preferably methyl or ethyl);    -   R⁸ is hydrogen or lower alkyl (preferably methyl or ethyl); and    -   each R¹⁰ is independently hydrogen or lower alkyl (preferably        methyl or ethyl), or two R¹⁰ together with the atom(s) attached        thereto form a 4- to 6-membered ring.

In some embodiments of the compound of Formula I-a or I-b

-   -   Q¹ and Q² are independently phenyl, 5-membered heteroaryl or        6-membered heteroaryl and are fused together in Formula I-a;    -   Q³ is 6-membered unsaturated ring wherein (1) the bond between        Y¹ and Y² is a double bond, and the bond between Y³ and Y² is a        single bond, or (2) the bond between    -   Y¹ and Y² is a single bond, and the bond between Y³ and Y² is a        double bond, and wherein Q³ is fused with Q² in Formula I-b;    -   s is 0 or 1; t is 1 or 2;    -   Y¹, Y³ and Y⁵ are independently C or N; Y², Y⁴ and Y⁶ are        independently CH, CR³ or N; provided that no more than four of        Y¹, Y², Y³, Y⁴, Y⁵ and Y⁶ are N;    -   R¹ is phenyl, 5-membered heteroaryl or 6-membered heteroaryl,        wherein the phenyl, 5-membered heteroaryl or 6-membered        heteroaryl is optionally substituted with 1 to 3 R⁶;    -   one of R and R⁴ is absent or is hydrogen, halo or lower alkyl        (preferably methyl or ethyl), and the other of R and R⁴ is L²-R⁵        or L³-R⁵;    -   X¹ is selected from the group consisting of OR⁸, NHR⁸, and SR⁸;    -   X² is selected from the group consisting of O, NR⁸, and S;    -   L¹ is selected from the group consisting —C(R⁸)₂—, —S—, —O— and        —NR⁸—;    -   L² is selected from the group consisting a covalent bond,        —C(O)N(R⁸)—, —N(R⁸)C(O)—, —N(R⁸)S(O)₂—, and —S(O)₂N(R⁸)—;    -   L³ is ═NC(O)—, or ═NS(O)₂—;    -   each R³ is independently selected from the group consisting of        lower alkyl (preferably methyl or ethyl) and halo;    -   R⁵ is phenyl, 5-membered heteroaryl, 6-membered heteroaryl,        5-membered heterocycloalkyl or 6-membered heterocycloalkyl;        wherein the phenyl, 5-membered heteroaryl or 6-membered        heteroaryl is optionally substituted with 1 to 4 R², wherein        each R² is independently selected from the group consisting of        lower alkyl, lower haloalkyl, —OH, —OR⁷, —SH, —SR⁷, —NR¹⁰R¹⁰,        halo, cyano, nitro, —COH, —COR⁷, —CO₂H, —CO₂R⁷, —CONR¹⁰R¹⁰,        —OCOR⁷, —OCO₂R⁷, —OCONR¹⁰R¹⁰, —NR¹⁰COR¹⁰, —NR¹⁰CO₂R¹⁰, —SOR⁷,        —SO₂R⁷, —SO₂NR¹⁰R¹⁰, and —NR¹⁰SO₂R⁷;    -   each R⁶ is independently selected from the group consisting of        halo and lower alkyl (preferably methyl or ethyl) optionally        substituted with 1-3 halo;    -   R⁷ is lower alkyl (preferably methyl or ethyl);    -   R⁸ is hydrogen or lower alkyl (preferably methyl or ethyl); and    -   each R¹⁰ is independently hydrogen or lower alkyl (preferably        methyl or ethyl), or two R¹⁰ together with the atom(s) attached        thereto form a 4- to 6-membered ring.

In one aspect, the present technology provides a method of treating acondition or disorder mediated by fascin activity in a subject in needthereof which method comprises administering to the subject atherapeutically effective amount of at least one compound, or acomposition comprising an effective amount of at least one compound, ofFormula I-c or I-d

or a tautomer thereof, and/or a pharmaceutically acceptable saltthereof, wherein

-   -   R¹ is phenyl, 5-membered heteroaryl or 6-membered heteroaryl;        wherein the phenyl, 5-membered heteroaryl or 6-membered        heteroaryl is optionally substituted with 1 to 3 R⁶;    -   R² is selected from the group consisting of lower alkyl, lower        haloalkyl, —OH, —OR⁷, —SH, —SR⁷, —NR¹⁰R¹⁰, halo, cyano, nitro,        —COH, —COR⁷, —CO₂H, —CO₂R⁷, —CONR¹⁰R¹⁰, —OCOR⁷, —OCO₂R⁷,        —OCONR¹⁰R¹⁰, —NR¹⁰COR¹⁰, —NR¹⁰CO₂R¹⁰, —SOR⁷, —SO₂R⁷,        —SO₂NR¹⁰R¹⁰, and —NR¹⁰SO₂R⁷;    -   each R³ is independently selected from the group consisting of        lower alkyl and halo;    -   m is 0, 1, 2 or 3;    -   n is 0, 1, 2, 3 or 4;    -   X¹ is selected from the group consisting of OR⁸, NHR⁸, and SR⁸;    -   X² is selected from the group consisting of O, NR⁸, and S;    -   L¹ is —S—, —O— or —NR⁸—;    -   L² is selected from the group consisting —C(O)N(R⁸)—,        —N(R⁸)C(O)—, —N(R⁸)S(O)₂—, and —S(O)₂N(R⁸)—;    -   L³ is ═NC(O)—, or ═NS(O)₂—;    -   each R⁶ is independently selected from the group consisting of        halo and lower alkyl optionally substituted with 1-3 halo;    -   R⁷ is lower alkyl;    -   R⁸ is hydrogen or lower alkyl; and    -   each R¹⁰ is independently hydrogen or lower alkyl, or two R¹⁰        together with the atom(s) attached thereto form a 4- to        6-membered ring.

In one embodiment, the present technology provides a method of treatinga condition or disorder mediated by fascin activity in a subject in needthereof which method comprises administering to the subject atherapeutically effective amount of at least one compound, or acomposition comprising an effective amount of at least one compound, ofFormula II

or a tautomer thereof, and/or a pharmaceutically acceptable saltthereof, wherein

-   -   ring A is a 5-membered heteroaryl or 5-membered        heterocycloalkyl;    -   W¹ and W⁴ are independently selected from the group consisting        of C, CR⁸, N, NR⁸, O and S, W² and W³ are independently C or N,        provided that at least one of W¹, W², W³, or W⁴ is C, and at        least one of W¹, W², W³, or W⁴ is N; wherein one of N is        optionally positively charged;    -   R²¹ and R²² are independently phenyl, 5-membered heteroaryl or        6-membered heteroaryl; wherein the phenyl, 5-membered heteroaryl        or 6-membered heteroaryl is optionally substituted with 1 to 3        R⁶;    -   R²³ is selected from the group consisting of hydrogen, lower        alkyl, phenyl, lower alkylphenyl, 5-membered heteroaryl and        6-membered heteroaryl; wherein the phenyl, lower alkylphenyl,        5-membered heteroaryl or 6-membered heteroaryl is optionally        substituted with 1 to 3 R⁶;    -   each R⁶ is independently selected from the group consisting of        halo and lower alkyl optionally substituted with 1-3 halo;    -   each R⁸ is independently hydrogen or lower alkyl; and    -   is a single or double bond, when        is a single bond, then R²⁴ is hydrogen or lower alkyl; when        is a double bond, then R²⁴ is absent.

In another embodiment, the present technology provides a method oftreating a condition or disorder mediated by fascin activity in asubject in need thereof which method comprises administering to thesubject a therapeutically effective amount of at least one compound, ora composition comprising an effective amount of at least one compound,of Formula III

or a tautomer, and/or a pharmaceutically acceptable salt thereof,wherein

-   -   R³⁰ is selected from the group consisting of lower alkyl, lower        alkenyl optionally substituted with phenyl, phenyl optionally        substituted with 1 or 2 substituents independently selected from        the group consisting of nitro and halo;    -   R³¹ is selected from the group consisting of lower haloalkyl,        —OH, —OR⁹, —SH, —SR⁷, —NR^(1I)R¹, halo, cyano, nitro, —COH,        —COR⁷, —CO₂H, —CO₂R⁷, —CONR¹⁰R¹⁰, —OCOR⁷, —OCO₂R⁷, —OCONR¹⁰R¹⁰,        —SO₂NR¹⁰R¹⁰, and —NR¹⁰SO₂R⁷;    -   p is 0, 1 or 2;    -   X³⁰ is C(═O) or S(O)₂;    -   R⁷ is lower alkyl;    -   R⁹ is phenyl; and    -   each R¹⁰ is independently hydrogen or lower alkyl, or two R⁰        together with the atom(s) attached thereto form a ring.

In one embodiment, the present technology provides a method ofinhibiting fascin activity, comprising administering an effective amountof a compound or a composition comprising an effective amount of acompound to a cell in need thereof to thereby inhibit fascin activity inthe cell, wherein the compound is of Formula I-a, I-b, II, or III, or atautomer, and/or a pharmaceutically acceptable salt thereof.

In another embodiment, the present technology provides a compound or acomposition comprising a compound for use in treating a condition ordisorder mediated by fascin activity in a subject in need thereof or ininhibiting fascin activity, wherein the compound is of Formula I-a, I-b,II, or III, or a tautomer, and/or a pharmaceutically acceptable saltthereof.

In another embodiment, the present technology provides use of a compoundor a composition comprising a compound in the preparation of amedicament for treating a condition or disorder mediated by fascinactivity in a subject in need thereof or for inhibiting fascin activity,wherein the compound is of Formula I-a, I-b, II, or III, or a tautomer,and/or a pharmaceutically acceptable salt thereof.

In some embodiments, the cell is in an animal. In some embodiments, thecell has been removed from an animal. In some embodiments, the animal isa human. In some embodiments, the human suffers from a disease orcondition.

In some embodiments, the condition or disorder is a metastatic cancer, aneuronal disorder, neuronal degeneration, an inflammatory condition, aviral infection, a bacterial infection, lymphoid hyperplasia, Hodgkin'sdisease or ischemia-related tissue damage. In some embodiments, thecondition or disorder is a metastatic cancer.

In some embodiments, the cancer is a carcinoma, lymphoma, sarcoma,melanoma, astrocytoma, mesothelioma cells, ovarian carcinoma, coloncarcinoma, pancreatic carcinoma, esophageal carcinoma, stomachcarcinoma, lung carcinoma, urinary carcinoma, bladder carcinoma, breastcancer, gastric cancer, leukemia, lung cancer, colon cancer, centralnervous system cancer, melanoma, ovarian cancer, renal cancer orprostate cancer.

BRIEF DESCRIPTION OF THE DRAWINGS

The invention will be more fully understood by reference to thefollowing drawings, which are for illustrative purposes only:

FIG. 1 illustrates the inhibition of breast tumor metastasis by2-chloro-N-(6-chlorobenzo[d]thiazol-2-yl)-5-nitrobenzenesulfonamide(Compound 3, NP-3), andN-(3-(1H-1,2,4-triazol-3-ylthio)-4-hydroxynaphthalen-1-yl)-4-methoxybenzenesulfonamide(Compound 10, NP-10) in mouse models. Lung metastasis was measured bythe 6-thioguanine clonogenic assay. Compound 3 (8 mg/kg) and Compound 10(30 mg/kg) were used. Results are mean±SD (n=5). *, P<0.01.

DETAILED DESCRIPTION

In the following detailed description, reference is made to theaccompanying drawings, which form a part hereof. In the drawings,similar symbols typically identify similar components, unless contextdictates otherwise. The illustrative embodiments described in thedetailed description, drawings, and claims are not meant to be limiting.Other embodiments may be utilized, and other changes may be made,without departing from the spirit or scope of the subject matterpresented here.

Fascin is an actin-bundling protein. For cell migration to proceed,actin cytoskeleton must be reorganized by forming polymers and bundlesto affect the dynamic changes of cell shapes (13-15). Individual actinfilaments are flexible and elongation of individual filaments per se isinsufficient for membrane protrusion which is necessary for cellmigration. Bundling of actin filaments provides rigidity to actinfilaments for protrusions in the form of lamellipodia and filopodiaagainst the compressive force from the plasma membrane (16) (17). Asnoted, one of the critical actin-bundling proteins is fascin (18-22).Fascin is the primary actin cross-linker in filopodia and shows nosequence homology with other actin-binding proteins (23). It is requiredto maximally cross-link the actin filaments into straight, compact, andrigid bundles (24).

Elevated levels of fascin have been found in many types of metastatictumors (including breast, prostate, ovarian, lung, gastric, esophageal,and others) and are correlated with clinically aggressive phenotypes,poor prognosis, and shorter survival (25-29) (30, 31) (32-34). Fascininhibitors may target tumor cell migration and invasion, and providetreatments for metastatic cancer.

Definitions

The technology is described herein using several definitions, as setforth throughout the specification.

The use of the terms “a” and “an” and “the” and similar referents in thecontext of describing the elements (especially in the context of thefollowing claims) are to be construed to cover both the singular and theplural, unless otherwise indicated herein or clearly contradicted bycontext.

As used herein, “about” will be understood by persons of ordinary skillin the art and will vary to some extent depending upon the context inwhich it is used. If there are uses of the term which are not clear topersons of ordinary skill in the art, given the context in which it isused, “about” will mean up to plus or minus 10% of the particular term.

A dash (“-”) that is not between two letters or symbols is used toindicate a point of attachment for a substituent. For example, —CONH₂ isattached through the carbon atom.

By “optional” or “optionally” is meant that the subsequently describedevent or circumstance may or may not occur, and that the descriptionincludes instances where the event or circumstance occurs and instancesin which it does not. For example, “optionally substituted alkyl”encompasses both “alkyl” and “substituted alkyl” as defined herein. Itwill be understood by those skilled in the art, with respect to anygroup containing one or more substituents, that such groups are notintended to introduce any substitution or substitution patterns that aresterically impractical, synthetically non-feasible and/or inherentlyunstable.

“Alkyl” encompasses straight chain and branched chain having theindicated number of carbon atoms, usually from 1 to 20 carbon atoms, forexample 1 to 8 carbon atoms, such as 1 to 6 carbon atoms. For exampleC₁-C₆ alkyl encompasses both straight and branched chain alkyl of from 1to 6 carbon atoms. Examples of alkyl groups include methyl, ethyl,propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, 2-pentyl,isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl, 3-methylpentyl, and thelike. Alkylene is another subset of alkyl, referring to the sameresidues as alkyl, but having two points of attachment. Alkylene groupswill usually have from 2 to 20 carbon atoms, for example 2 to 8 carbonatoms, such as from 2 to 6 carbon atoms. For example, C₀ alkyleneindicates a covalent bond and C₁ alkylene is a methylene group. When analkyl residue having a specific number of carbons is named, allgeometric isomers having that number of carbons are intended to beencompassed; thus, for example, “butyl” is meant to include n-butyl,sec-butyl, isobutyl and t-butyl; “propyl” includes n-propyl andisopropyl. “Lower alkyl” refers to an alkyl group having 1 to 4 carbons.

“Alkenyl” refers to straight or branched hydrocarbyl groups having theindicated number of carbon atoms, usually from 1 to 8 carbon atoms, forexample 2 to 4 carbon atoms, and at least 1 and preferably from 1 to 2sites of vinyl (>C═C<) unsaturation. Such groups are exemplified, forexample, by vinyl, allyl, and but-3-en-1-yl. Included within this termare the cis and trans isomers or mixtures of these isomers. “Loweralkenyl” refers to an alkenyl group having 1 to 4 carbons, which can beindicated by C₂-C₄ alkenyl.

“Cycloalkyl” indicates a non-aromatic, fully saturated carbocyclic ringhaving the indicated number of carbon atoms, for example, 3 to 10, or 3to 8, or 3 to 6 ring carbon atoms. Cycloalkyl groups may be monocyclicor polycyclic (e.g., bicyclic, tricyclic). Examples of cycloalkyl groupsinclude cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl andcyclohexyl, as well as bridged and caged ring groups (e.g., norbornane,bicyclo[2.2.2]octane). In addition, one ring of a polycyclic cycloalkylgroup may be aromatic, provided the polycyclic cycloalkyl group is boundto the parent structure via a non-aromatic carbon. For example, a1,2,3,4-tetrahydronaphthalen-1-yl group (wherein the moiety is bound tothe parent structure via a non-aromatic carbon atom) is a cycloalkylgroup, while 1,2,3,4-tetrahydronaphthalen-5-yl (wherein the moiety isbound to the parent structure via an aromatic carbon atom) is notconsidered a cycloalkyl group. Examples of polycyclic cycloalkyl groupsconsisting of a cycloalkyl group fused to an aromatic ring are describedbelow.

“Aryl” indicates an aromatic carbon ring having the indicated number ofcarbon atoms, for example, 6 to 12 or 6 to 10 carbon atoms. Aryl groupsmay be monocyclic or polycyclic (e.g., bicyclic, tricyclic). In someinstances, both rings of a polycyclic aryl group are aromatic (e.g.,naphthyl). In other instances, polycyclic aryl groups may include anon-aromatic ring (e.g., cycloalkyl, cycloalkenyl, heterocycloalkyl,heterocycloalkenyl) fused to an aromatic ring, provided the polycyclicaryl group is bound to the parent structure via an atom in the aromaticring. Thus, a 1,2,3,4-tetrahydronaphthalen-5-yl group (wherein themoiety is bound to the parent structure via an aromatic carbon atom) isconsidered an aryl group, while 1,2,3,4-tetrahydronaphthalen-1-yl(wherein the moiety is bound to the parent structure via a non-aromaticcarbon atom) is not considered an aryl group. Similarly, a1,2,3,4-tetrahydroquinolin-8-yl group (wherein the moiety is bound tothe parent structure via an aromatic carbon atom) is considered an arylgroup, while 1,2,3,4-tetrahydroquinolin-1-yl group (wherein the moietyis bound to the parent structure via a non-aromatic nitrogen atom) isnot considered an aryl group. However, the term “aryl” does notencompass or overlap with “heteroaryl”, as defined herein, regardless ofthe point of attachment (e.g., both quinolin-5-yl and quinolin-2-yl areheteroaryl groups). In some instances, aryl is phenyl or naphthyl. Incertain instances, aryl is phenyl. Additional examples of aryl groupscomprising an aromatic carbon ring fused to a non-aromatic ring aredescribed below.

“Carboxy” or “carboxyl” refers to —COOH or a salt thereof.

“Heteroaryl” indicates an aromatic ring containing the indicated numberof atoms (e.g., 5 to 12, or 5 to 10 membered heteroaryl) made up of oneor more heteroatoms (e.g., 1, 2, 3 or 4 heteroatoms) selected from N, Oand S and with the remaining ring atoms being carbon. 5-Memberedheteroaryl is a heteroaryl having 5 ring atoms. 6-Membered heteroaryl isa heteroaryl having 6 ring atoms. Heteroaryl groups do not containadjacent S and O atoms. In some embodiments, the total number of S and Oatoms in the heteroaryl group is not more than 2. In some embodiments,the total number of S and O atoms in the heteroaryl group is not morethan 1. Unless otherwise indicated, heteroaryl groups may be bound tothe parent structure by a carbon or nitrogen atom, as valency permits.For example, “pyridyl” includes 2-pyridyl, 3-pyridyl and 4-pyridylgroups, and “pyrrolyl” includes 1-pyrrolyl, 2-pyrrolyl and 3-pyrrolylgroups. When nitrogen is present in a heteroaryl ring, it may, where thenature of the adjacent atoms and groups permits, exist in an oxidizedstate (i.e., N⁺—O⁻). Additionally, when sulfur is present in aheteroaryl ring, it may, where the nature of the adjacent atoms andgroups permits, exist in an oxidized state (i.e., S⁺—O⁻ or SO₂).Heteroaryl groups may be monocyclic or polycyclic (e.g., bicyclic,tricyclic).

In some instances, a heteroaryl group is monocyclic. Examples includepyrrole, pyrazole, imidazole, triazole (e.g., 1,2,3-triazole,1,2,4-triazole, 1,2,4-triazole), tetrazole, furan, isoxazole, oxazole,oxadiazole (e.g., 1,2,3-oxadiazole, 1,2,4-oxadiazole, 1,3,4-oxadiazole),thiophene, isothiazole, thiazole, thiadiazole (e.g., 1,2,3-thiadiazole,1,2,4-thiadiazole, 1,3,4-thiadiazole), pyridine, pyridazine, pyrimidine,pyrazine, triazine (e.g., 1,2,4-triazine, 1,3,5-triazine) and tetrazine.

In some instances, both rings of a polycyclic heteroaryl group arearomatic. Examples include indole, isoindole, indazole, benzoimidazole,benzotriazole, benzofuran, benzoxazole, benzoisoxazole, benzoxadiazole,benzothiophene, benzothiazole, benzoisothiazole, benzothiadiazole,1H-pyrrolo[2,3-b]pyridine, 1H-pyrazolo[3,4-b]pyridine,3H-imidazo[4,5-b]pyridine, 3H-[1,2,3]triazolo[4,5-b]pyridine,1H-pyrrolo[3,2-b]pyridine, 1H-pyrazolo[4,3-b]pyridine,1H-imidazo[4,5-b]pyridine, 1H-[1,2,3]triazolo[4,5-b]pyridine,1H-pyrrolo[2,3-c]pyridine, 1H-pyrazolo[3,4-c]pyridine,3H-imidazo[4,5-c]pyridine, 3H-[1,2,3]triazolo[4,5-c]pyridine,1H-pyrrolo[3,2-c]pyridine, 1H-pyrazolo[4,3-c]pyridine,1H-imidazo[4,5-c]pyridine, 1H-[1,2,3]triazolo[4,5-c]pyridine,furo[2,3-b]pyridine, oxazolo[5,4-b]pyridine, isoxazolo[5,4-b]pyridine,[1,2,3]oxadiazolo[5,4-b]pyridine, furo[3,2-b]pyridine,oxazolo[4,5-b]pyridine, isoxazolo[4,5-b]pyridine,[1,2,3]oxadiazolo[4,5-b]pyridine, furo[2,3-c]pyridine,oxazolo[5,4-c]pyridine, isoxazolo[5,4-c]pyridine,[1,2,3]oxadiazolo[5,4-c]pyridine, furo[3,2-c]pyridine,oxazolo[4,5-c]pyridine, isoxazolo[4,5-c]pyridine,[1,2,3]oxadiazolo[4,5-c]pyridine, thieno[2,3-b]pyridine,thiazolo[5,4-b]pyridine, isothiazolo[5,4-b]pyridine,[1,2,3]thiadiazolo[5,4-b]pyridine, thieno[3,2-b]pyridine,thiazolo[4,5-b]pyridine, isothiazolo[4,5-b]pyridine,[1,2,3]thiadiazolo[4,5-b]pyridine, thieno[2,3-c]pyridine,thiazolo[5,4-c]pyridine, isothiazolo[5,4-c]pyridine,[1,2,3]thiadiazolo[5,4-c]pyridine, thieno[3,2-c]pyridine,thiazolo[4,5-c]pyridine, isothiazolo[4,5-c]pyridine,[1,2,3]thiadiazolo[4,5-c]pyridine, quinoline, isoquinoline, cinnoline,quinazoline, quinoxaline, phthalazine, naphthyridine (e.g.,1,8-naphthyridine, 1,7-naphthyridine, 1,6-naphthyridine,1,5-naphthyridine, 2,7-naphthyridine, 2,6-naphthyridine),imidazo[1,2-a]pyridine, 1H-pyrazolo[3,4-d]thiazole,1H-pyrazolo[4,3-d]thiazole and imidazo[2,1-b]thiazole.

In other instances, polycyclic heteroaryl groups may include anon-aromatic ring (e.g., cycloalkyl, cycloalkenyl, heterocycloalkyl,heterocycloalkenyl) fused to a heteroaryl ring, provided the polycyclicheteroaryl group is bound to the parent structure via an atom in thearomatic ring. For example, a 4,5,6,7-tetrahydrobenzo[d]thiazol-2-ylgroup (wherein the moiety is bound to the parent structure via anaromatic carbon atom) is considered a heteroaryl group, while4,5,6,7-tetrahydrobenzo[d]thiazol-5-yl (wherein the moiety is bound tothe parent structure via a non-aromatic carbon atom) is not considered aheteroaryl group. Examples of polycyclic heteroaryl groups consisting ofa heteroaryl ring fused to a non-aromatic ring are described below.

“Heterocycloalkyl” indicates a non-aromatic, fully saturated ring havingthe indicated number of atoms (e.g., 3 to 10, or 3 to 7, memberedheterocycloalkyl) made up of one or more heteroatoms (e.g., 1, 2, 3 or 4heteroatoms) selected from N, O and S and with the remaining ring atomsbeing carbon. 5-Membered heterocycloalkyl is a heterocycloalkyl having 5ring atoms. 6-Membered heterocycloalkyl is a heterocycloalkyl having 6ring atoms. Heterocycloalkyl groups may be monocyclic or polycyclic(e.g., bicyclic, tricyclic). Examples of heterocycloalkyl groups includeoxiranyl, aziridinyl, azetidinyl, pyrrolidinyl, imidazolidinyl,pyrazolidinyl, piperidinyl, piperazinyl, morpholinyl andthiomorpholinyl. When nitrogen is present in a heterocycloalkyl ring, itmay, where the nature of the adjacent atoms and groups permits, exist inan oxidized state (i.e., N⁺—O⁻). Examples include piperidinyl N-oxideand morpholinyl-N-oxide. Additionally, when sulfur is present in aheterocycloalkyl ring, it may, where the nature of the adjacent atomsand groups permits, exist in an oxidized state (i.e., S⁺—O⁻ or —SO₂—).Examples include thiomorpholine S-oxide and thiomorpholine S,S-dioxide.In addition, one ring of a polycyclic heterocycloalkyl group may bearomatic (e.g., aryl or heteroaryl), provided the polycyclicheterocycloalkyl group is bound to the parent structure via anon-aromatic carbon or nitrogen atom. For example, a1,2,3,4-tetrahydroquinolin-1-yl group (wherein the moiety is bound tothe parent structure via a non-aromatic nitrogen atom) is considered aheterocycloalkyl group, while 1,2,3,4-tetrahydroquinolin-8-yl group(wherein the moiety is bound to the parent structure via an aromaticcarbon atom) is not considered a heterocycloalkyl group. Examples ofpolycyclic heterocycloalkyl groups consisting of a heterocycloalkylgroup fused to an aromatic ring are described below.

By “alkoxy” is meant an alkyl group of the indicated number of carbonatoms attached through an oxygen bridge such as, for example, methoxy,ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, pentoxy,2-pentyloxy, isopentoxy, neopentoxy, hexoxy, 2-hexoxy, 3-hexoxy,3-methylpentoxy, and the like. An alkoxy group is further meant toencompass a cycloalkyl group, as defined above, that is likewiseattached through an oxygen bridge. Alkoxy groups will usually have from1 to 6 carbon atoms attached through the oxygen bridge. “Lower alkoxy”refers to an alkoxy group having 1 to 4 carbons.

The term “halo” includes fluoro, chloro, bromo, and iodo, and the term“halogen” includes fluorine, chlorine, bromine, and iodine.

The term “substituted”, as used herein, means that any one or morehydrogens on the designated atom or group is replaced with a selectionfrom the indicated group, provided that the designated atom's normalvalence is not exceeded. When a substituent is oxo (i.e., ═O) then 2hydrogens on the atom are replaced. Combinations of substituents and/orvariables are permissible only if such combinations result in stablecompounds or useful synthetic intermediates. A stable compound or stablestructure is meant to imply a compound that is sufficiently robust tosurvive isolation from a reaction mixture, and subsequent formulation asan agent having at least practical utility. Unless otherwise specified,substituents are named into the core structure. For example, it is to beunderstood that when (cycloalkyl)alkyl is listed as a possiblesubstituent, the point of attachment of this substituent to the corestructure is in the alkyl portion.

“Haloalkyl” refers to alkyl groups substituted with 1 to 5, 1 to 3, or 1to 2 halo groups, wherein alkyl and halo are as defined herein.

“Lower alkylphenyl” refers to C₁-C₄ alkyl-phenyl.

“Isomers” are different compounds that have the same molecular formula.“Stereoisomers” are isomers that differ only in the way the atoms arearranged in space. “Enantiomers” are stereoisomers that arenon-superimposable mirror images of each other. A 1:1 mixture of a pairof enantiomers is a “racemic” mixture. The symbol “(+)” may be used todesignate a racemic mixture where appropriate. “Diastereoisomers” arestereoisomers that have at least two asymmetric atoms, but which are notmirror-images of each other. A “meso compound” or “meso isomer” is anon-optically active member of a set of stereoisomers. Meso isomerscontain two or more stereocenters but are not chiral (i.e., a plane ofsymmetry exists within the molecule). The absolute stereochemistry isspecified according to the Cahn-Ingold-Prelog R—S system. When acompound is a pure enantiomer the stereochemistry at each chiral carboncan be specified by either R or S. Resolved compounds whose absoluteconfiguration is unknown can be designated (+) or (−) depending on thedirection (dextro- or levorotatory) which they rotate plane polarizedlight at the wavelength of the sodium D line. Certain of the compoundsdisclosed and/or described herein contain one or more asymmetric centersand can thus give rise to enantiomers, diastereomers, meso isomers andother stereoisomeric forms. Unless otherwise indicated, compoundsdisclosed and/or described herein include all such possible enantiomers,diastereomers, meso isomers and other stereoisomeric forms, includingracemic mixtures, optically pure forms and intermediate mixtures.Enantiomers, diastereomers, meso isomers and other stereoisomeric formscan be prepared using chiral synthons or chiral reagents, or resolvedusing conventional techniques. Unless specified otherwise, when thecompounds disclosed and/or described herein contain olefinic doublebonds or other centers of geometric asymmetry, it is intended that thecompounds include both E and Z isomers.

“Tautomers” are structurally distinct isomers that interconvert bytautomerization. Tautomerization is a form of isomerization and includesprototropic or proton-shift tautomerization, which is considered asubset of acid-base chemistry. Prototropic tautomerization orproton-shift tautomerization involves the migration of a protonaccompanied by changes in bond order, often the interchange of a singlebond with an adjacent double bond. Where tautomerization is possible(e.g. in solution), a chemical equilibrium of tautomers can be reached.An example of tautomerization is keto-enol tautomerization. A specificexample of keto-enol tautomerization is the interconverision ofpentane-2,4-dione and 4-hydroxypent-3-en-2-one tautomers. Anotherexample of tautomerization is phenol-keto tautomerization. A specificexample of phenol-keto tautomerization is the interconversion ofpyridin-4-ol and pyridin-4(1H)-one tautomers. When the compoundsdescribed herein contain moieties capable of tautomerization, and unlessspecified otherwise, it is intended that the compounds include allpossible tautomers.

Pharmaceutically acceptable forms of the compounds recited hereininclude pharmaceutically acceptable salts, and mixtures thereof.

“Pharmaceutically acceptable salts” include, but are not limited tosalts with inorganic acids, such as hydrochlorate, phosphate,diphosphate, hydrobromate, sulfate, sulfinate, nitrate, and like salts;as well as salts with an organic acid, such as malate, maleate,fumarate, tartrate, succinate, citrate, acetate, lactate,methanesulfonate, p-toluenesulfonate, 2-hydroxyethylsulfonate, benzoate,salicylate, stearate, and alkanoate such as acetate, HOOC—(CH₂)_(n)—COOHwhere n is 0-4, and like salts. Similarly, pharmaceutically acceptablecations include, but are not limited to sodium, potassium, calcium,aluminum, lithium, and ammonium.

In addition, if the compounds described herein are obtained as an acidaddition salt, the free base can be obtained by basifying a solution ofthe acid salt. Conversely, if the product is a free base, an additionsalt, particularly a pharmaceutically acceptable addition salt, may beproduced by dissolving the free base in a suitable organic solvent andtreating the solution with an acid, in accordance with conventionalprocedures for preparing acid addition salts from base compounds. Thoseskilled in the art will recognize various synthetic methodologies thatmay be used to prepare non-toxic pharmaceutically acceptable additionsalts.

The compounds disclosed and/or described herein can be enriched isotopicforms, e.g., enriched in the content of ²H, ³H, ¹¹C, ¹³C and/or ¹⁴C. Inone embodiment, the compound contains at least one deuterium atom. Suchdeuterated forms can be made, for example, by the procedure described inU.S. Pat. Nos. 5,846,514 and 6,334,997. Such deuterated compounds mayimprove the efficacy and increase the duration of action of compoundsdisclosed and/or described herein. Deuterium substituted compounds canbe synthesized using various methods, such as those described in: Dean,D., Recent Advances in the Synthesis and Applications of RadiolabeledCompounds for Drug Discovery and Development, Curr. Pharm. Des., 2000;6(10); Kabalka, G. et al., The Synthesis of Radiolabeled Compounds viaOrganometallic Intermediates, Tetrahedron, 1989, 45(21), 6601-21; andEvans, E., Synthesis of radiolabeled compounds, J. Radioanal. Chem.,1981, 64(1-2), 9-32.

As used herein the terms “group”, “radical” or “fragment” are synonymousand are intended to indicate functional groups or fragments of moleculesattachable to a bond or other fragments of molecules.

The term “active agent” is used to indicate a substance which hasbiological activity. In some embodiments, an “active agent” is asubstance having pharmaceutical utility. For example an active agent maybe an anti-metastasis therapeutic.

The term “therapeutically effective amount” means an amount effective,when administered to a human or non-human subject, to provide atherapeutic benefit such as amelioration of symptoms, slowing of diseaseprogression, or prevention of disease e.g., a therapeutically effectiveamount may be an amount sufficient to decrease the symptoms of a diseaseresponsive to inhibition of fascin activity.

“Inhibition of fascin activity” refers to a decrease in fascin activityas a direct or indirect response to the presence of at least onecompound, or pharmaceutically acceptable salt thereof, described herein,relative to the activity of fascin in the absence of the at least onecompound, or pharmaceutically acceptable salt thereof, described herein.The decrease in activity may be due to the direct interaction of the atleast one compound, or pharmaceutically acceptable salt thereof,described herein with fascin or with one or more other factors that inturn affect fascin activity.

In some embodiments, the compound, or pharmaceutically acceptable saltthereof, described herein has an IC₅₀ (the concentration that inhibits50% of fascin activity) value of about 500 micromolar, about 100micromolar, about 10 micromolar, about 1 micromolar, about 500nanomolar, about 400 nanomolar, about 300 nanomolar, about 200nanomolar, about 100 nanomolar, about 50 nanomolar, about 10 nanomolar,of less than about 10 nanomolar, or a range between and including anytwo of these values.

A “disease responsive to inhibition of fascin activity” is a disease inwhich inhibiting fascin provides a therapeutic benefit such as anamelioration of symptoms, decrease in disease progression, prevention ordelay of disease onset, prevention or amelioration of an inflammatoryresponse, or inhibition of aberrant activity and/or death of certaincell-types (such as cancer cells).

“Treatment” or “treating” means any treatment of a disease in a patient,including:

-   -   a) preventing the disease, that is, causing the clinical        symptoms of the disease not to develop;    -   b) inhibiting the progression of the disease;    -   c) slowing or arresting the development of clinical symptoms;        and/or    -   d) relieving the disease, that is, causing the regression of        clinical symptoms.

“Subject” or “patient” refers to an animal, such as a mammal, that hasbeen or will be the object of treatment, observation or experiment. Themethods described herein may be useful in both human therapy andveterinary applications. In some embodiments, the subject is a mammal;and in some embodiments the subject is human.

As used herein, the term “cancer” includes solid mammalian tumors aswell as hematological malignancies. The terms “tumor cell(s)” and“cancer cell(s)” are used interchangeably herein.

“Solid mammalian tumors” include cancers of the head and neck, lung,mesothelioma, mediastinum, esophagus, stomach, pancreas, hepatobiliarysystem, small intestine, colon, colorectal, rectum, anus, kidney,urethra, bladder, prostate, urethra, penis, testis, gynecologicalorgans, ovaries, breast, endocrine system, skin, central nervous system;sarcomas of the soft tissue and bone; and melanoma of cutaneous andintraocular origin.

The term “hematological malignancies” includes childhood leukemia andlymphomas, Hodgkin's disease, lymphomas of lymphocytic and cutaneousorigin, acute and chronic leukemia, plasma cell neoplasm and cancersassociated with AIDS.

Also, in these examples and elsewhere, abbreviations have the followingmeanings:

-   -   ° C.=degree Celsius    -   μL=microliter    -   M=micromolar    -   DDT=dithiothreitol    -   DMSO=dimethyl sulfoxide    -   g=gram    -   kg=kilogram    -   hr or h=hour    -   L=liter    -   M=molar    -   nM=nanomolar    -   mg=milligram    -   MHz=mega Hertz    -   min=minute    -   mL=milliliter    -   mM=millimolar    -   mmol=millimole    -   mol=mole    -   PMSF=phenylmethylsulfonyl fluoride    -   N=normal    -   EDTA=ethylenediaminetetraacetic acid    -   μm=micrometer    -   r.p.m=round per minute    -   S.D.=standard deviation    -   v/v=volume/volume    -   wt=weight        Methods of Treatment

In one aspect, the present technology provides a method of treating acondition or disorder mediated by fascin activity in a subject in needthereof which method comprises administering to the subject atherapeutically effective amount of at least one compound of Formula I-aor I-b

or a tautomer thereof, and/or a pharmaceutically acceptable saltthereof, wherein

-   -   Q¹ and Q² are independently phenyl, 5-membered heteroaryl or        6-membered heteroaryl and are fused together in Formula I-a;    -   Q³ is unsaturated ring wherein (1) the bond between Y¹ and Y² is        a double bond, and the bond between Y³ and Y² is a single bond,        or (2) the bond between Y¹ and Y² is a single bond, and the bond        between Y³ and Y² is a double bond, and wherein Q³ is fused with        Q² in Formula I-b;    -   s is 0 or 1; t is 1 or 2;    -   Y¹, Y³ and Y⁵ are independently C or N; Y², Y⁴ and Y⁶ are        independently CH, CR³ or N; provided that no more than four of        Y¹, Y², Y³, Y⁴, Y⁵ and Y⁶ are N;    -   R¹ is phenyl, 5-membered heteroaryl or 6-membered heteroaryl,        wherein the phenyl, 5-membered heteroaryl or 6-membered        heteroaryl is optionally substituted with 1 to 3 R⁶;    -   one of R and R⁴ is absent or is hydrogen, halo or lower alkyl        (preferably methyl or ethyl), and the other of R and R⁴ is L²-R⁵        or L³-R⁵; or R is absent and R⁴ is —(CH₂)_(j)—R¹¹; j is 1, 2 or        3; R¹¹ is selected from the group consisting of —OH, —OR⁷, —SH,        —SR⁷, —NR¹⁰R¹⁰, cyano, nitro, —COH, —COR⁷, —CO₂H, —CO₂R⁷,        —CONR¹⁰R¹⁰, —OCOR⁷, —OCO₂R⁷, —OCONR¹⁰R¹⁰, —NR¹⁰COR¹⁰,        —NR¹⁰CO₂R¹⁰, —SOR⁷, —SO₂R⁷, —SO₂NR¹⁰R¹⁰, and —NR¹⁰SO₂R⁷;    -   X¹ is selected from the group consisting of OR⁸, NHR⁸, and SR⁸;    -   X² is selected from the group consisting of O, NR⁸, and S;    -   L¹ is selected from the group consisting —(C(R⁸)₂)_(j)—,        —(C(R⁸)₂)_(q)—C(O)—(C(R⁸)₂)_(r)—,        —(C(R⁸)₂)_(q)—C(O)N(R⁸)—(C(R⁸)₂)_(r)—,        —(C(R⁸)₂)_(q)—N(R⁸)C(O)—(C(R⁸)₂)_(r)—,        —(C(R⁸)₂)_(q)—N(R⁸)S(O)₂—(C(R⁸)₂)_(r)—,        —(CH₂)_(q)—S(O)₂N(R⁸)—(CH₂)_(r)—, —S—, —O— and —NR⁸—;    -   q is 0 or 1;    -   r is 0 or 1;    -   L² is selected from the group consisting a covalent bond,        —C(O)N(R⁸)—, —N(R⁸)C(O)—, —N(R⁸)S(O)₂—, and —S(O)₂N(R⁸)—;    -   L³ is ═NC(O)—, or ═NS(O)₂—;    -   each R³ is independently selected from the group consisting of        lower alkyl (preferably methyl or ethyl) and halo;    -   R⁵ is phenyl, 5-membered heteroaryl, 6-membered heteroaryl,        5-membered heterocycloalkyl or 6-membered heterocycloalkyl;        wherein the phenyl, 5-membered heteroaryl or 6-membered        heteroaryl is optionally substituted with 1 to 4 R², wherein        each R² is independently selected from the group consisting of        lower alkyl, lower haloalkyl, —OH, —OR⁷, —SH, —SR⁷, —NR¹⁰R¹⁰,        halo, cyano, nitro, —COH, —COR⁷, —CO₂H, —CO₂R⁷, —CONR¹⁰R¹⁰,        —OCOR⁷, —OCO₂R⁷, —OCONR¹⁰R¹⁰, —NR¹⁰COR¹⁰, —NR¹⁰CO₂R¹⁰, —SOR⁷,        —SO₂R⁷, —SO₂NR¹⁰R¹⁰, and —NR¹⁰SO₂R⁷;    -   each R⁶ is independently selected from the group consisting of        halo and lower alkyl (preferably methyl or ethyl) optionally        substituted with 1-3 halo; or two adjacent R⁶ on a phenyl ring        form a 5- or 6-membered cycloalkyl or heterocycloalkyl fused        with the phenyl ring;    -   R⁷ is lower alkyl (preferably methyl or ethyl);    -   R⁸ is hydrogen or lower alkyl (preferably methyl or ethyl); and    -   each R¹⁰ is independently hydrogen or lower alkyl (preferably        methyl or ethyl), or two R¹⁰ together with the atom(s) attached        thereto form a 4- to 6-membered heterocycloalkyl ring.

In some embodiments, provided is a method of treating a condition ordisorder mediated by fascin activity in a subject in need thereof whichmethod comprises administering to the subject a therapeuticallyeffective amount of at least one compound of Formula I-c or I-d

or a tautomer thereof, and/or a pharmaceutically acceptable saltthereof, wherein

-   -   R¹ is phenyl, 5-membered heteroaryl or 6-membered heteroaryl;        wherein the phenyl, 5-membered heteroaryl or 6-membered        heteroaryl is optionally substituted with 1 to 3 R⁶;    -   R² is selected from the group consisting of lower alkyl, lower        haloalkyl, —OH, —OR⁷, —SH, —SR⁷, —NR¹⁰R¹⁰, halo, cyano, nitro,        —COH, —COR⁷, —CO₂H, —CO₂R⁷, —CONR¹⁰R¹⁰, —OCOR⁷, —OCO₂R⁷,        —OCONR¹⁰R¹⁰, —NR¹⁰COR¹⁰, —NR¹⁰CO₂R¹⁰, —SOR⁷, —SO₂R⁷,        —SO₂NR¹⁰R¹⁰, and —NR¹⁰SO₂R⁷;    -   each R³ is independently selected from the group consisting of        lower alkyl and halo;    -   m is 0, 1, 2 or 3;    -   n is 0, 1, 2, 3 or 4;    -   X¹ is selected from the group consisting of OR⁸, NHR⁸, and SR⁸;    -   X² is selected from the group consisting of O, NR⁸, and S;    -   L¹ is —S—, —O— or —NR⁸—;    -   L² is selected from the group consisting —C(O)N(R⁸)—,        —N(R⁸)C(O)—, —N(R⁸)S(O)₂—, and —S(O)₂N(R⁸)—;    -   L³ is ═NC(O)— or ═NS(O)₂—;    -   each R⁶ is independently selected from the group consisting of        halo and lower alkyl optionally substituted with 1-3 halo;    -   R⁷ is lower alkyl;    -   R⁸ is hydrogen or lower alkyl; and    -   each R¹⁰ is independently hydrogen or lower alkyl, or two R¹⁰        together with the atom(s) attached thereto form a 4- to        6-membered ring.

In some embodiments of Formula I-b, Q is 6-membered unsaturated ring ands is 1.

In some embodiments, L¹ is O. In some embodiments, L¹ is S. In someembodiments, L¹ is —NH—. In some embodiments, L¹ is —NCH₃—.

In some embodiments, L² is —N(R⁸)S(O)₂—. In some embodiments, L² is—NHS(O)₂—. In some embodiments, L³ is ═NS(O)₂—.

In some embodiments, the compound is of Formula I-e or I-f

or a tautomer, and/or pharmaceutically acceptable salt thereof.

In some embodiments, the compound is of Formula I-g or I-h

or a tautomer, and/or pharmaceutically acceptable salt thereof.

In some embodiments, the compound is of Formula I-i or I-j

or a tautomer, and/or pharmaceutically acceptable salt thereof,wherein Y is N or CR, R is hydrogen or lower alkyl, and R¹, L¹, L², andR⁵ are as defined in Formula I-a or I-b.

In some embodiments, the compound is of Formula I-k

or a tautomer, and/or pharmaceutically acceptable salt thereof,wherein R¹, L¹ and R⁵ are as defined in Formula I-b.

In some embodiments, the compound is of Formula I-1 or I-m

wherein L¹, R², and R⁶ are as defined in Formula I-b, n is 0, 1, 2, 3 or4 and u is 1, 2 or 3.

In some embodiments, the compound is of Formula I-n

or a tautomer, and/or pharmaceutically acceptable salt thereof,wherein R¹, L¹, j and R¹¹ are as defined in Formula I-b. In someembodiments, R¹¹ is OH.

In some embodiments, R¹ is phenyl. In some embodiments, R¹ is phenylsubstituted with 1 to 3 R⁶. In some embodiments, R¹ is phenylsubstituted with one group chosen from halo and lower alkyl optionallysubstituted with 1-3 halo. In some embodiments, R¹ is phenyl substitutedwith two groups chosen from halo and lower alkyl optionally substitutedwith 1-3 halo. In some embodiments, R¹ is phenyl substituted with threegroups chosen from halo and lower alkyl optionally substituted with 1-3halo. In some embodiments, R¹ is trifluoromethylphenyl. In someembodiments, R¹ is dichlorophenyl. In some embodiments, R¹ is phenylsubstituted with two adjacent R⁶ that form a 5- or 6-membered cycloalkylfused with the phenyl ring. In some embodiments, R¹ is phenylsubstituted with two adjacent R⁶ that form a 5- or 6-memberedheterocycloalkyl (such as a heterocycloalkyl comprising one or two ringoxygen atoms) fused with the phenyl ring.

In some embodiments, R¹ is unsubstituted 5-membered heteroaryl. In someembodiments, R¹ is 5-membered heteroaryl substituted with one groupchosen from halo and lower alkyl. In some embodiments, R¹ is 5-memberedheteroaryl substituted with two groups chosen from halo and lower alkyl.In some embodiments, R¹ is 5-membered heteroaryl substituted with threegroups chosen from halo and lower alkyl. In some embodiments, R¹ isunsubstituted 6-membered heteroaryl. In some embodiments, R¹ is6-membered heteroaryl substituted with one group chosen from halo andlower alkyl. In some embodiments, R¹ is 6-membered heteroarylsubstituted with two groups chosen from halo and lower alkyl. In someembodiments, R¹ is 6-membered heteroaryl substituted with three groupschosen from halo and lower alkyl.

In some embodiments, R¹ is unsubstituted triazole. In some embodiments,R¹ is triazole substituted with one group chosen from halo and loweralkyl.

In some embodiments, X¹ is OH. In some embodiments, X² is O.

In some embodiments, m is 0. In some embodiments, m is 1.

In some embodiments, R³ is halo. In some embodiments, R³ is lower alkyl.

In some embodiments, n is 1. In some embodiments, n is 2. In someembodiments, n is 3.

In some embodiments, R² is independently selected from the groupconsisting of OH, halo, lower alkyl, and —OR⁷. In some embodiments, R²is selected from the group consisting of bromo, methyl, ethyl, methoxy,and ethoxy. In some embodiments, R² is halo. In some embodiments, R² is—OR⁷. In some embodiments, R² is methyl. In some embodiments, R² isethyl. In some embodiment, n is 2 or 3, and each R² is methyl.

In some embodiments, L¹ is —(C(R⁸)₂)_(j)—,—(C(R⁸)₂)_(q)—C(O)—(C(R⁸)₂)_(r)—, —(C(R⁸)₂)_(q)—C(O)N(R⁸)—(C(R⁸)₂)_(r)—,—(C(R⁸)₂)_(q)—N(R⁸)C(O)—(C(R⁸)₂)_(r)—,—(C(R⁸)₂)_(q)—N(R⁸)S(O)₂—(C(R⁸)₂)_(r)—, or—(CH₂)_(q)—S(O)₂N(R⁸)—(CH₂)_(r)—. In some embodiments, R⁸ is hydrogen.In some embodiments, j is 1. In some embodiments, L¹ is —(CH₂)_(j)—. Insome embodiments, L¹ is methylene. In some embodiments, L¹ is —CH₂C(O)—.In some embodiments, L¹ is —C(O)CH₂—. In some embodiments, L¹ is—CH₂—C(O)NH—CH₂—. In some embodiments, L¹ is —CH₂—NHC(O)—CH₂—. In someembodiments, L¹ is —NHC(O)—CH₂—. In some embodiments, L¹ is—CH₂—NHC(O)—. In some embodiments, L¹ is —C(O)NH—CH₂—. In someembodiments, L¹ is —CH₂—C(O)NH—. In some embodiments, L¹ is selectedfrom the group consisting —S—, —O— and —NR⁸—. In some embodiments, L¹ is—S—. In some embodiments, L¹ is —O—. In some embodiments, L¹ is —NR⁸—.

In some embodiments, Y is N.

In some embodiments, L² is a covalent bond and R⁵ is a 5-memberedheterocycloalkyl or 6-membered heterocycloalkyl. In some embodiments,the 5-membered heterocycloalkyl or 6-membered heterocycloalkyl comprisesa sulfur ring atom which is oxidized to SO₂. In some embodiments, L² is—NHC(O)— and R⁵ is a 5-membered heteroaryl or 6-membered heteroaryl.

In some embodiments, L¹ is methylene. In some embodiments, R¹ is phenyl.

In some embodiments, the compound is selected from

-   N-(3-(1H-1,2,4-triazol-3-ylthio)-4-hydroxynaphthalen-1-yl)-2,5-dimethylbenzenesulfonamide;-   N-(3-(1H-1,2,4-triazol-3-ylthio)-4-hydroxynaphthalen-1-yl)-4-ethoxybenzenesulfonamide;-   N-(3-(1H-1,2,4-triazol-3-ylthio)-4-hydroxynaphthalen-1-yl)-4-methoxybenzenesulfonamide;-   N-(3-(1H-1,2,4-triazol-3-ylthio)-4-hydroxynaphthalen-1-yl)-4-ethylbenzenesulfonamide;-   N-(3-(1H-1,2,4-triazol-3-ylthio)-4-hydroxynaphthalen-1-yl)-2,4,5-trimethylbenzenesulfonamide;-   (Z)—N-(3-(1H-1,2,4-triazol-3-ylthio)-4-oxonaphthalen-1(4H)-ylidene)benzenesulfonamide;-   N-(3-(1H-1,2,4-triazol-3-ylthio)-4-hydroxynaphthalen-1-yl)-4-bromobenzenesulfonamide;    and-   N-(3-(1H-1,2,4-triazol-3-ylthio)-4-hydroxynaphthalen-1-yl)-2,4-dimethylbenzenesulfonamide;-   or a tautomer, and/or pharmaceutically acceptable salt thereof.

In some embodiments, the compound is

-   5-(3,4-dichlorobenzyl)-1-(S,S-dioxo-tetrahydrothiophen-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one    or-   N-(1-(4-(trifluoromethyl)benzyl)-1H-indazol-3-yl)furan-2-carboxamide,-   or a tautomer, and/or pharmaceutically acceptable salt thereof.

In some embodiments, the compound is selected from the group consistingof

-   5-(3-chlorobenzyl)-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one;-   2-(4-oxo-1-(S,S-dioxo-tetrahydrothiophen-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-5(4H)-yl)-N-(3-(trifluoromethyl)phenyl)acetamide;-   N-(4-fluorobenzyl)-2-(4-oxo-1-(S,    S-dioxo-tetrahydrothiophen-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-5    (4H)-yl)acetamide;-   N-(benzo[d][1,3]dioxol-5-ylmethyl)-2-(4-oxo-1-(S,S-dioxo-tetrahydrothiophen-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-5(4H)-yl)acetamide;-   N-(4-chlorophenyl)-2-(4-oxo-1-(S,    S-dioxo-tetrahydrothiophen-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-5    (4H)-yl)acetamide;-   5-(2-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-2-oxoethyl)-1-(S,    S-dioxo-tetrahydrothiophen-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one;-   5-(2-(2,4-dimethylphenyl)-2-oxoethyl)-1-(S,    S-dioxo-tetrahydrothiophen-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one;-   5-(2-(benzo[d][1,3]dioxol-5-yl)-2-oxoethyl)-1-(S,S-dioxo-tetrahydrothiophen-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one;-   5-(3,4-dichlorobenzyl)-1-o-tolyl-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one;    5-(3,4-dichlorobenzyl)-1-(2,3-dimethylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one;    and-   5-(3,4-dichlorobenzyl)-1-(2,4-dimethylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one;-   or a tautomer, and/or pharmaceutically acceptable salt thereof.

Also provided is a method of treating a condition or disorder mediatedby fascin activity in a subject in need thereof which method comprisesadministering to the subject a therapeutically effective amount of atleast one compound of Formula II

or a tautomer thereof, and/or a pharmaceutically acceptable salt thereofwherein

-   -   ring A is a 5-membered heteroaryl or 5-membered        heterocycloalkyl;    -   W¹ and W⁴ are independently selected from the group consisting        of C, CR⁸, N, NR⁸, O and S, W² and W³ are independently C or N,        provided that at least one of W¹, W², W³, or W⁴ is C, and at        least one of W¹, W², W³, or W⁴ is N; wherein one of N is        optionally positively charged;    -   R²¹ and R²² are independently phenyl, 5-membered heteroaryl or        6-membered heteroaryl; wherein the phenyl, 5-membered heteroaryl        or 6-membered heteroaryl is optionally substituted with 1 to 3        R⁶;    -   R²³ is selected from the group consisting of hydrogen, lower        alkyl, phenyl, lower alkylphenyl, 5-membered heteroaryl and        6-membered heteroaryl; wherein the phenyl, lower alkylphenyl,        5-membered heteroaryl or 6-membered heteroaryl is optionally        substituted with 1 to 3 R⁶;    -   each R⁶ is independently selected from the group consisting of        halo and lower alkyl optionally substituted with 1-3 halo;    -   is a single or double bond, when        is a single bond, then R²⁴ is hydrogen or lower alkyl; when        is a double bond, then R²⁴ is absent.

In some embodiments, ring A is thiadiazole.

In some embodiments, the compound is of Formula II-a or II-b

or a tautomer thereof, and/or a pharmaceutically acceptable saltthereof.

In some embodiments, W⁴ is S. In some embodiments, W⁴ is O. In someembodiments, W⁴ is NH.

In some embodiments, R²¹ and R²² are, independently, phenyl optionallysubstituted with halo or alkyl. In some embodiments, R²¹ and R²² arephenyl. In some embodiments, R²¹ and R²² are, independently, phenylsubstituted with one, two, or three groups chosen from halo and loweralkyl. In some embodiments, R²¹ and R²² are, independently, phenylsubstituted with one group chosen from halo and lower alkyl. In someembodiments, R²¹ and R²² are, independently, phenyl substituted with twogroups chosen from halo and lower alkyl. In some embodiments, R²¹ andR²² are, independently, phenyl substituted with three groups chosen fromhalo and lower alkyl.

In some embodiments, R²³ is methyl, phenyl, or benzyl. In someembodiments, R²³ is methyl. In some embodiments, R²³ is phenyl. In someembodiments, R²³ is benzyl. In some embodiments, R²³ is loweralkylphenyl. In some embodiments, R²³ is 5-membered heteroaryl or6-membered heteroaryl.

In some embodiments, R²⁴ is hydrogen.

In some embodiments, the compound is selected from

-   (Z)—N-(2,3-diphenyl-1,2,4-thiadiazol-5 (2H)-ylidene)methanamine;-   N-methyl-2,3-diphenyl-1,2,4-thiadiazolium-5-amine;-   N-benzyl-2,3-diphenyl-1,2,4-thiadiazolium-5-amine; and-   N-phenyl-2,3-diphenyl-1,2,4-thiadiazolium-5-amine;-   or a tautomer, and/or pharmaceutically acceptable salt thereof.

In another embodiment, the present technology provides a method oftreating a condition or disorder mediated by fascin activity in asubject in need thereof which method comprises administering to thesubject a therapeutically effective amount of at least one compound ofFormula III

or a tautomer, and/or a pharmaceutically acceptable salt thereof,wherein

-   -   R³⁰ is selected from the group consisting of lower alkyl, lower        alkenyl optionally substituted with phenyl, phenyl optionally        substituted with 1 or 2 substituents independently selected from        the group consisting of nitro and halo;    -   R³¹ is selected from the group consisting of lower haloalkyl,        —OH, —OR⁹, —SH, —SR⁷, —NR¹⁰R¹⁰, halo, cyano, nitro, —COH, —COR⁷,        —CO₂H, —CO₂R⁷, —CONR¹⁰R¹⁰, —OCOR⁷, —OCO₂R⁷, —OCONR¹⁰R¹⁰,        —SO₂NR¹⁰R¹⁰, and —NR¹⁰SO₂R⁷;    -   p is 0, 1 or 2;    -   X³⁰ is C(═O) or S(O)₂;    -   R⁷ is lower alkyl;    -   R⁹ is phenyl; and    -   each R¹⁰ is independently hydrogen or lower alkyl, or two R¹⁰        together with the atom(s) attached thereto form a ring.

In some embodiments, R³⁰ is lower alkyl.

In some embodiments, R³⁰ is lower alkenyl. In some embodiments, R³⁰ islower alkenyl substituted with phenyl.

In some embodiments, R³⁰ is phenyl optionally substituted with one ortwo substituents selected from the group consisting of nitro and halo,for example, chloro, bromo or fluoro.

In some embodiments, X³⁰ is C(═O) and R³⁰ is lower alkyl. In someembodiments, X³⁰ is C(═O) and R³⁰ is lower alkenyl optionallysubstituted with phenyl.

In some embodiments, X³⁰ is S(O)₂ and R³⁰ is phenyl. In someembodiments, X³⁰ is S(O)₂ and R³⁰ is phenyl substituted with one or twosubstituents independently nitro or halo, for example, chloro.

In some embodiments, R³¹ is selected from the group consisting of haloand phenoxy. In some embodiments, R³¹ is selected from the groupconsisting of fluoro, chloro and phenoxy.

In some embodiments, p is 0. In some embodiments, p is 1. In someembodiments, p is 2.

In some embodiments, the compound is selected from the group consistingof

-   2-chloro-N-(6-chlorobenzo[d]thiazol-2-yl)-5-nitrobenzenesulfonamide,-   3-chloro-N-(6-phenoxybenzo[d]thiazol-2-yl)benzenesulfonamide,-   N-(6-fluorobenzo[d]thiazol-2-yl)-3-nitrobenzenesulfonamide,-   2,3-dichloro-N-(6-fluorobenzo[d]thiazol-2-yl)benzenesulfonamide,-   N-(6-chlorobenzo[d]thiazol-2-yl)acetamide, and-   N-(benzo[d]thiazol-2-yl)cinnamamide,-   or a tautomer, and/or pharmaceutically acceptable salt thereof.

In one embodiment the present technology provides a method of inhibitingfascin activity, comprising administering an effective amount of afascin inhibitor to a cell to thereby inhibit fascin activity in thecell, wherein the fascin inhibitor is of Formula I-a, I-b, II, or III.In some embodiments, the fascin inhibitor has a fascin inhibition IC₅₀of no more than 100 μM. In some embodiments, the fascin inhibitor has afascin inhibition IC₅₀ of no more than 50 μM. In some embodiments, thefascin inhibitor has a fascin inhibition IC₅₀ of no more than 20 μM. Insome embodiments, the fascin inhibitor has a fascin inhibition IC₅₀ ofno more than 8 μM.

In some embodiments, the condition or disorder is a metastatic cancer, aneuronal disorder, neuronal degeneration, an inflammatory condition, aviral infection, a bacterial infection, lymphoid hyperplasia, Hodgkin'sdisease or ischemia-related tissue damage.

In some embodiments, the condition or disorder is a metastatic cancer.

In some embodiments, the cancer is a carcinoma, lymphoma, sarcoma,melanoma, astrocytoma, mesothelioma cells, ovarian carcinoma, coloncarcinoma, pancreatic carcinoma, esophageal carcinoma, stomachcarcinoma, lung carcinoma, urinary carcinoma, bladder carcinoma, breastcancer, gastric cancer, leukemia, lung cancer, colon cancer, centralnervous system cancer, melanoma, ovarian cancer, renal cancer orprostate cancer. In some embodiments, the cancer is lung cancer, breastcancer or prostate cancer.

In another aspect, the present technology provides is a method ofinhibiting fascin activity, comprising administering an effective amountof a fascin inhibitor to a cell to thereby inhibit fascin activity inthe cell, wherein the fascin inhibitor is of Formula I-a or I-b

or a tautomer thereof, and/or a pharmaceutically acceptable saltthereof, wherein

-   -   Q¹ and Q² are independently phenyl, 5-membered heteroaryl or        6-membered heteroaryl and are fused together in Formula I-a;    -   Q³ is unsaturated ring wherein (1) the bond between Y¹ and Y² is        a double bond, and the bond between Y³ and Y² is a single bond,        or (2) the bond between Y¹ and Y² is a single bond, and the bond        between Y³ and Y² is a double bond, and wherein Q³ is fused with        Q² in Formula I-b;    -   s is 0 or 1; t is 1 or 2;    -   Y¹, Y³ and Y⁵ are independently C or N; Y², Y⁴ and Y⁶ are        independently CH, CR³ or N; provided that no more than four of        Y¹, Y², Y³, Y⁴, Y⁵ and Y⁶ are N;    -   R¹ is phenyl, 5-membered heteroaryl or 6-membered heteroaryl,        wherein the phenyl, 5-membered heteroaryl or 6-membered        heteroaryl is optionally substituted with 1 to 3 R⁶;    -   one of R and R⁴ is absent or is hydrogen, halo or lower alkyl        (preferably methyl or ethyl), and the other of R and R⁴ is L²-R⁵        or L³-R⁵; or R is absent and R⁴ is —(CH₂)_(j)—R¹¹; j is 1, 2 or        3; R¹¹ is selected from the group consisting of —OH, —OR⁷, —SH,        —SR⁷, —NR¹⁰R¹⁰, cyano, nitro, —COH, —COR⁷, —CO₂H, —CO₂R⁷,        —CONR¹⁰R¹⁰, —OCOR⁷, —OCO₂R⁷, —OCONR¹⁰R¹⁰, —NR¹⁰COR¹⁰,        —NR¹⁰CO₂R¹⁰, —SOR⁷, —SO₂R⁷, —SO₂NR¹⁰R¹⁰, and —NR¹⁰SO₂R⁷;    -   X¹ is selected from the group consisting of OR⁸, NHR⁸, and SR⁸;    -   X² is selected from the group consisting of O, NR⁸, and S;    -   L¹ is selected from the group consisting —(C(R⁸)₂)_(j)—,        —(C(R⁸)₂)_(q)—C(O)—(C(R⁸)₂)_(r)—,        —(C(R⁸)₂)_(q)—C(O)N(R⁸)—(C(R⁸)₂)_(r)—,        —(C(R⁸)₂)_(q)—N(R⁸)C(O)—(C(R⁸)₂)_(r)—,        —(C(R⁸)₂)_(q)—N(R⁸)S(O)₂—(C(R⁸)₂)_(r)—,        —(CH₂)_(q)—S(O)₂N(R⁸)—(CH₂)_(r)—, —S—, —O— and —NR⁸—;    -   q is 0 or 1;    -   r is 0 or 1;    -   L² is selected from the group consisting a covalent bond,        —C(O)N(R⁸)—, —N(R⁸)C(O)—, —N(R⁸)S(O)₂—, and —S(O)₂N(R⁸)—;    -   L³ is ═NC(O)—, or ═NS(O)₂—;    -   each R³ is independently selected from the group consisting of        lower alkyl (preferably methyl or ethyl) and halo;    -   R⁵ is phenyl, 5-membered heteroaryl, 6-membered heteroaryl,        5-membered heterocycloalkyl or 6-membered heterocycloalkyl;        wherein the phenyl, 5-membered heteroaryl or 6-membered        heteroaryl is optionally substituted with 1 to 4 R², wherein        each R² is independently selected from the group consisting of        lower alkyl, lower haloalkyl, —OH, —OR⁷, —SH, —SR⁷, —NR¹⁰R¹⁰,        halo, cyano, nitro, —COH, —COR⁷, —CO₂H, —CO₂R⁷, —CONR¹⁰R¹⁰,        —OCOR⁷, —OCO₂R⁷, —OCONR¹⁰R¹⁰, —NR¹⁰COR¹⁰, —NR¹⁰CO₂R¹⁰, —SOR⁷,        —SO₂R⁷, —SO₂NR¹⁰R¹⁰, and —NR¹⁰SO₂R⁷;    -   each R⁶ is independently selected from the group consisting of        halo and lower alkyl (preferably methyl or ethyl) optionally        substituted with 1-3 halo; or two adjacent R⁶ on a phenyl ring        form a 5- or 6-membered cycloalkyl or heterocycloalkyl fused        with the phenyl ring;    -   R⁷ is lower alkyl (preferably methyl or ethyl);    -   R⁸ is hydrogen or lower alkyl (preferably methyl or ethyl); and    -   each R¹⁰ is independently hydrogen or lower alkyl (preferably        methyl or ethyl), or two R¹⁰ together with the atom(s) attached        thereto form a 4- to 6-membered heterocycloalkyl ring.

In some embodiments of Formula I-b, Q is 6-membered unsaturated ring ands is 1.

In some embodiments, L¹ is O. In some embodiments, L¹ is S. In someembodiments, L¹ is —NH—. In some embodiments, L¹ is —NCH₃—. In someembodiments, L¹ is —CH₂—.

In some embodiments, L² is —N(R⁸)S(O)₂—. In some embodiments, L² is—NHC(O)—. In some embodiments, L² is —NHS(O)₂—. In some embodiments, L²is a covalent bond. In some embodiments, L² is —C(O)NH—. In someembodiments, L³ is ═NS(O)₂—.

In some embodiments, provided is a method of inhibiting fascin activity,comprising administering an effective amount of a fascin inhibitor to acell to thereby inhibit fascin activity in the cell, wherein the fascininhibitor is of Formula I-c or I-d

or a tautomer thereof, and/or a pharmaceutically acceptable saltthereof, wherein

-   -   R¹ is phenyl, 5-membered heteroaryl or 6-membered heteroaryl;        wherein the phenyl, 5-membered heteroaryl or 6-membered        heteroaryl is optionally substituted with 1 to 3 R⁶;    -   R² is selected from the group consisting of lower alkyl, lower        haloalkyl, —OH, —OR⁷, —SH, —SR⁷, —NR¹⁰R¹⁰, halo, cyano, nitro,        —COH, —COR⁷, —CO₂H, —CO₂R⁷, —CONR¹⁰R¹⁰, —OCOR⁷, —OCO₂R⁷,        —OCONR¹⁰R¹⁰, —NR¹⁰COR¹⁰, —NR¹⁰CO₂R¹⁰, —SOR⁷, —SO₂R⁷,        —SO₂NR¹⁰R¹⁰, and —NR¹⁰SO₂R⁷;    -   each R³ is independently selected from the group consisting of        lower alkyl and halo;    -   m is 0, 1, 2 or 3;    -   n is 0, 1, 2, 3 or 4;    -   X¹ is selected from the group consisting of OR⁸, NHR⁸, and SR⁸;    -   X² is selected from the group consisting of O, NR⁸, and S;    -   L¹ is —S—, —O— or —NR⁸—;    -   L² is selected from the group consisting —C(O)N(R⁸)—,        —N(R⁸)C(O)—, —N(R⁸)S(O)₂—, and —S(O)₂N(R⁸)—;    -   L³ is ═NC(O)— or ═NS(O)₂—;    -   each R⁶ is independently selected from the group consisting of        halo and lower alkyl optionally substituted with 1-3 halo;    -   R⁷ is lower alkyl;    -   R⁸ is hydrogen or lower alkyl; and    -   each R¹⁰ is independently hydrogen or lower alkyl, or two R¹⁰        together with the atom(s) attached thereto form a 4- to        6-membered ring.

In some embodiments, L¹ is O. In some embodiments, L¹ is S. In someembodiments, L¹ is —NH—. In some embodiments, L¹ is —NCH₃—.

In some embodiments, L² is —N(R⁸)S(O)₂—. In some embodiments, L² is—NHS(O)₂—. In some embodiments, L³ is ═NS(O)₂—.

In some embodiments, the compound is of Formula I-e or I-f

or a tautomer thereof, and/or a pharmaceutically acceptable saltthereof.

In some embodiments, the compound is of Formula I-g or I-h

or a tautomer thereof, and/or a pharmaceutically acceptable saltthereof, wherein “

” represents that the single bond can be on either side of the doublebond.

In some embodiments, the compound is of Formula I-i or I-j

or a tautomer, and/or pharmaceutically acceptable salt thereof,wherein Y is N or CR, R is hydrogen or lower alkyl, and R¹, L¹, L², andR⁵ are as defined in Formula I-a or I-b.

In some embodiments, the compound is of Formula I-k

or a tautomer, and/or pharmaceutically acceptable salt thereof,wherein R¹, L¹ and R⁵ are as defined in Formula I-b.

In some embodiments, the compound is of Formula I-1 or I-m

wherein L¹, R², R⁶ and n are as defined in Formula I-b and u is 1, 2 or3.

In some embodiments, the compound is of Formula I-n

or a tautomer, and/or pharmaceutically acceptable salt thereof,wherein R¹, L¹, j and R¹¹ are as defined in Formula I-b. In someembodiments, R¹¹ is OH.

In some embodiments, R¹ is phenyl. In some embodiments, R¹ is phenylsubstituted with 1 to 3 R⁶. In some embodiments, R¹ is phenylsubstituted with two adjacent R⁶ that form a 5- or 6-membered cycloalkylfused with the phenyl ring. In some embodiments, R¹ is phenylsubstituted with two adjacent R⁶ that form a 5- or 6-memberedheterocycloalkyl (such as a heterocycloalkyl comprising one or two ringoxygen atoms) fused with the phenyl ring. In some embodiments, R¹ isphenyl substituted with one group chosen from halo and lower alkyloptionally substituted with 1-3 halo. In some embodiments, R¹ is phenylsubstituted with two groups chosen from halo and lower alkyl optionallysubstituted with 1-3 halo. In some embodiments, R¹ is phenyl substitutedwith three groups chosen from halo and lower alkyl optionallysubstituted with 1-3 halo. In some embodiments, R¹ istrifluoromethylphenyl. In some embodiments, R¹ is dichlorophenyl.

In some embodiments, R¹ is unsubstituted 5-membered heteroaryl. In someembodiments, R¹ is 5-membered heteroaryl substituted with one groupchosen from halo and lower alkyl. In some embodiments, R¹ is 5-memberedheteroaryl substituted with two groups chosen from halo and lower alkyl.In some embodiments, R¹ is 5-membered heteroaryl substituted with threegroups chosen from halo and lower alkyl. In some embodiments, R¹ isunsubstituted 6-membered heteroaryl. In some embodiments, R¹ is6-membered heteroaryl substituted with one group chosen from halo andlower alkyl. In some embodiments, R¹ is 6-membered heteroarylsubstituted with two groups chosen from halo and lower alkyl. In someembodiments, R¹ is 6-membered heteroaryl substituted with three groupschosen from halo and lower alkyl.

In some embodiments, R¹ is unsubstituted triazole. In some embodiments,R¹ is triazole substituted with one group chosen from halo and loweralkyl.

In some embodiments, X¹ is OH. In some embodiments, X² is O.

In some embodiments, m is 0. In some embodiments, m is 1.

In some embodiments, R³ is halo. In some embodiments, R³ is lower alkyl.

In some embodiments, n is 1. In some embodiments, n is 2. In someembodiments, n is 3.

In some embodiments, R² is independently selected from the groupconsisting of OH, halo, lower alkyl, and —OR⁷. In some embodiments, R²is selected from the group consisting of bromo, methyl, ethyl, methoxy,and ethoxy. In some embodiments, R² is halo. In some embodiments, R² is—OR⁷. In some embodiments, R² is methyl. In some embodiments, R² isethyl. In some embodiment, n is 2 or 3, and each R² is methyl.

In some embodiments, L¹ is —(C(R⁸)₂)_(j)—,—(C(R⁸)₂)_(q)—C(O)—(C(R⁸)₂)_(r)—, —(C(R⁸)₂)_(q)—C(O)N(R⁸)—(C(R⁸)₂)_(r)—,—(C(R⁸)₂)_(q)—N(R⁸)C(O)—(C(R⁸)₂)_(r)—,—(C(R⁸)₂)_(q)—N(R⁸)S(O)₂—(C(R⁸)₂)_(r)—, or—(CH₂)_(q)—S(O)₂N(R⁸)—(CH₂)_(r)—. In some embodiments, R⁸ is hydrogen.In some embodiments, j is 1. In some embodiments, L¹ is —(CH₂)_(j)—. Insome embodiments, L¹ is methylene. In some embodiments, L¹ is —CH₂C(O)—.In some embodiments, L¹ is —C(O)CH₂—. In some embodiments, L¹ is—CH₂—C(O)NH—CH₂—. In some embodiments, L¹ is —CH₂—NHC(O)—CH₂—. In someembodiments, L¹ is —NHC(O)—CH₂—. In some embodiments, L¹ is—CH₂—NHC(O)—. In some embodiments, L¹ is —C(O)NH—CH₂—. In someembodiments, L¹ is —CH₂—C(O)NH—. In some embodiments, L¹ is selectedfrom the group consisting —S—, —O— and —NR⁸—. In some embodiments, L¹ is—S—. In some embodiments, L¹ is —O—. In some embodiments, L¹ is —NR⁸—.

In some embodiments, Y is N.

In some embodiments, L² is a covalent bond and R⁵ is a 5-memberedheterocycloalkyl or 6-membered heterocycloalkyl. In some embodiments,the 5-membered heterocycloalkyl or 6-membered heterocycloalkyl comprisesa sulfur ring atom which is oxidized to SO₂. In some embodiments, L² is—NHC(O)— and R⁵ is a 5-membered heteroaryl or 6-membered heteroaryl. Insome embodiments, L¹ is methylene. In some embodiments, R¹ is phenyl.

In some embodiments, the compound is selected from

-   N-(3-(1H-1,2,4-triazol-3-ylthio)-4-hydroxynaphthalen-1-yl)-2,5-dimethylbenzenesulfonamide;-   N-(3-(1H-1,2,4-triazol-3-ylthio)-4-hydroxynaphthalen-1-yl)-4-ethoxybenzenesulfonamide;-   N-(3-(1H-1,2,4-triazol-3-ylthio)-4-hydroxynaphthalen-1-yl)-4-methoxybenzenesulfonamide;-   N-(3-(1H-1,2,4-triazol-3-ylthio)-4-hydroxynaphthalen-1-yl)-4-ethylbenzenesulfonamide;-   N-(3-(1H-1,2,4-triazol-3-ylthio)-4-hydroxynaphthalen-1-yl)-2,4,5-trimethylbenzenesulfonamide;-   (Z)—N-(3-(1H-1,2,4-triazol-3-ylthio)-4-oxonaphthalen-1(4H)-ylidene)benzenesulfonamide;-   N-(3-(1H-1,2,4-triazol-3-ylthio)-4-hydroxynaphthalen-1-yl)-4-bromobenzenesulfonamide;    and-   N-(3-(1H-1,2,4-triazol-3-ylthio)-4-hydroxynaphthalen-1-yl)-2,4-dimethylbenzenesulfonamide;-   or a tautomer, and/or pharmaceutically acceptable salt thereof.

In some embodiments, the compound is

-   5-(3,4-dichlorobenzyl)-1-(S,S-dioxo-tetrahydrothiophen-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one    or-   N-(1-(4-(trifluoromethyl)benzyl)-1H-indazol-3-yl)furan-2-carboxamide,-   or a tautomer, and/or pharmaceutically acceptable salt thereof.

In some embodiments, the compound is selected from the group consistingof

-   5-(3-chlorobenzyl)-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one;-   2-(4-oxo-1-(S,S-dioxo-tetrahydrothiophen-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-5(4H)-yl)-N-(3-(trifluoromethyl)phenyl)acetamide;-   N-(4-fluorobenzyl)-2-(4-oxo-1-(S,    S-dioxo-tetrahydrothiophen-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-5    (4H)-yl)acetamide;-   N-(benzo[d][1,3]dioxol-5-ylmethyl)-2-(4-oxo-1-(S,S-dioxo-tetrahydrothiophen-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-5(4H)-yl)acetamide;-   N-(4-chlorophenyl)-2-(4-oxo-1-(S,    S-dioxo-tetrahydrothiophen-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-5    (4H)-yl)acetamide;-   5-(2-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-2-oxoethyl)-1-(S,    S-dioxo-tetrahydrothiophen-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one;-   5-(2-(2,4-dimethylphenyl)-2-oxoethyl)-1-(S,    S-dioxo-tetrahydrothiophen-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one;-   5-(2-(benzo[d][1,3]dioxol-5-yl)-2-oxoethyl)-1-(S,S-dioxo-tetrahydrothiophen-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one;-   5-(3,4-dichlorobenzyl)-1-o-tolyl-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one;-   5-(3,4-dichlorobenzyl)-1-(2,3-dimethylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one;    and-   5-(3,4-dichlorobenzyl)-1-(2,4-dimethylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one;-   or a tautomer, and/or pharmaceutically acceptable salt thereof.

Also provided is a method of inhibiting fascin activity, comprisingadministering an effective amount of a fascin inhibitor to a cell tothereby inhibit fascin activity in the cell, wherein the fascininhibitor is a compound of Formula II

or a tautomer thereof, and/or a pharmaceutically acceptable salt thereofwherein

-   -   ring A is a 5-membered heteroaryl or 5-membered        heterocycloalkyl;    -   W¹ and W⁴ are independently selected from the group consisting        of C, CR⁸, N, NR⁸, O and S, W² and W³ are independently C or N,        provided that at least one of W¹, W², W³, or W⁴ is C, and at        least one of W¹, W², W³, or W⁴ is N; wherein one of N is        optionally positively charged;    -   R²¹ and R²² are independently phenyl, 5-membered heteroaryl or        6-membered heteroaryl; wherein the phenyl, 5-membered heteroaryl        or 6-membered heteroaryl is optionally substituted with 1 to 3        R⁶;    -   R²³ is selected from the group consisting of hydrogen, lower        alkyl, phenyl, lower alkylphenyl, 5-membered heteroaryl and        6-membered heteroaryl; wherein the phenyl, lower alkylphenyl,        5-membered heteroaryl or 6-membered heteroaryl is optionally        substituted with 1 to 3 R⁶;    -   each R⁶ is independently selected from the group consisting of        halo and lower alkyl optionally substituted with 1-3 halo;    -   each R⁸ is independently hydrogen or lower alkyl; and    -   is a single or double bond, when        is a single bond, then R²⁴ is hydrogen or lower alkyl; when        is a double bond, then R²⁴ is absent.

In some embodiments, ring A is thiadiazole.

In some embodiments, the method comprises a compound of Formula II-a orII-b

or a tautomer thereof, and/or a pharmaceutically acceptable saltthereof.

In some embodiments, W⁴ is S. In some embodiments, W⁴ is O. In someembodiments, W⁴ is NH.

In some embodiments, R²¹ and R²² are, independently, phenyl optionallysubstituted with halo or alkyl. In some embodiments, R²¹ and R²² arephenyl. In some embodiments, R²¹ and R²² are, independently, phenylsubstituted with one, two, or three groups chosen from halo and loweralkyl. In some embodiments, R²¹ and R²² are, independently, phenylsubstituted with one group chosen from halo and lower alkyl. In someembodiments, R²¹ and R²² are, independently, phenyl substituted with twogroups chosen from halo and lower alkyl. In some embodiments, R²¹ andR²² are, independently, phenyl substituted with three groups chosen fromhalo and lower alkyl.

In some embodiments, R²³ is methyl, phenyl, or benzyl. In someembodiments, R²³ is methyl. In some embodiments, R²³ is phenyl. In someembodiments, R²³ is benzyl. In some embodiments, R²³ is alkylsubstituted with a phenyl. In some embodiments, R²³ is alkyl substituteda 5- or 6-membered heteroaryl.

In some embodiments, R²⁴ is hydrogen.

In some embodiments, the compound is selected from

-   (Z)—N-(2,3-diphenyl-1,2,4-thiadiazol-5 (2H)-ylidene)methanamine;-   N-methyl-2,3-diphenyl-1,2,4-thiadiazolium-5-amine;-   N-benzyl-2,3-diphenyl-1,2,4-thiadiazolium-5-amine; and-   N-phenyl-2,3-diphenyl-1,2,4-thiadiazolium-5-amine;-   or a tautomer, and/or pharmaceutically acceptable salt thereof.

Also provided is a method of inhibiting fascin activity, comprisingadministering an effective amount of a fascin inhibitor to a cell tothereby inhibit fascin activity in the cell, wherein the fascininhibitor is of Formula III

or a tautomer, and/or a pharmaceutically acceptable salt thereof,wherein

-   -   R³⁰ is selected from the group consisting of lower alkyl, lower        alkenyl optionally substituted with phenyl, phenyl optionally        substituted with 1 or 2 substituents independently selected from        the group consisting of nitro and halo;    -   R³¹ is selected from the group consisting of lower haloalkyl,        —OH, —OR⁹, —SH, —SR⁷, —NR¹⁰R¹⁰, halo, cyano, nitro, —COH, —COR⁷,        —CO₂H, —CO₂R⁷, —CONR¹⁰R¹⁰, —OCOR⁷, —OCO₂R⁷, —OCONR¹⁰R¹⁰,        —SO₂NR¹⁰R¹⁰, and —NR¹⁰SO₂R⁷;    -   p is 0, 1 or 2;    -   X³⁰ is C(═O) or S(O)₂;    -   R⁷ is lower alkyl;    -   R⁹ is phenyl; and    -   each R¹⁰ is independently hydrogen or lower alkyl, or two R¹⁰        together with the atom(s) attached thereto form a ring.

In some embodiments, R³⁰ is lower alkyl.

In some embodiments, R³⁰ is lower alkenyl. In some embodiments, R³⁰ islower alkenyl substituted with phenyl.

In some embodiments, R³⁰ is phenyl optionally substituted with one ortwo substituents selected from the group consisting of nitro and halo,for example, chloro, bromo or fluoro.

In some embodiments, X³⁰ is C(═O) and R³⁰ is lower alkyl. In someembodiments, X³⁰ is C(═O) and R³⁰ is lower alkenyl optionallysubstituted with phenyl.

In some embodiments, X³⁰ is S(O)₂ and R³⁰ is phenyl. In someembodiments, X³⁰ is S(O)₂ and R³⁰ is phenyl substituted with one or twosubstituents independently nitro or halo, for example, chloro.

In some embodiments, R³¹ is selected from the group consisting of haloand phenoxy. In some embodiments, R³¹ is selected from the groupconsisting of fluoro, chloro, and phenoxy.

In some embodiments, p is 0. In some embodiments, p is 1. In someembodiments, p is 2.

In some embodiments, the compound is selected from the group consistingof

-   2-chloro-N-(6-chlorobenzo[d]thiazol-2-yl)-5-nitrobenzenesulfonamide,-   3-chloro-N-(6-phenoxybenzo[d]thiazol-2-yl)benzenesulfonamide,-   N-(6-fluorobenzo[d]thiazol-2-yl)-3-nitrobenzenesulfonamide,-   2,3-dichloro-N-(6-fluorobenzo[d]thiazol-2-yl)benzenesulfonamide,-   N-(6-fluorobenzo[d]thiazol-2-yl)acetamide,-   N-(6-chlorobenzo[d]thiazol-2-yl)acetamide and-   N-(benzo[d]thiazol-2-yl)cinnamamide,-   or a tautomer, and/or pharmaceutically acceptable salt thereof.

In some embodiments, the cell is in an animal. In some embodiments, thecell has been removed from an animal. In some embodiments, the animal isa human. In some embodiments, the human suffers from a disease orcondition.

In some embodiments, the condition or disorder is a metastatic cancer, aneuronal disorder, neuronal degeneration, an inflammatory condition, aviral infection, a bacterial infection, lymphoid hyperplasia, Hodgkin'sdisease or ischemia-related tissue damage. In some embodiments, thecondition or disorder is a metastatic cancer.

In some embodiments, the cancer is a carcinoma, lymphoma, sarcoma,melanoma, astrocytoma, mesothelioma cells, ovarian carcinoma, coloncarcinoma, pancreatic carcinoma, esophageal carcinoma, stomachcarcinoma, lung carcinoma, urinary carcinoma, bladder carcinoma, breastcancer, gastric cancer, leukemia, lung cancer, colon cancer, centralnervous system cancer, melanoma, ovarian cancer, renal cancer orprostate cancer. In some embodiments, the cancer is lung cancer, breastcancer, or prostate cancer.

Agents that modulate the activity of fascin can be used to treat avariety of diseases and conditions. For example, as illustrated herein,fascin promotes actin bundling and plays a key role in cell migrationand metastasis of cancer cells. Hence, modulators and inhibitors offascin can be used to treat and inhibit metastatic cancer.

However, fascin also plays a role in other diseases and conditions. Forexample, neurite shape and trajectory is modulated by fascin (Kraft etal., Phenotypes of Drosophila brain neurons in primary culture reveal arole for fascin in neurite shape and trajectory, J. Neurosci,26(34):8734-47 (2006)). Fascin is also involved in neuronal degeneration(Fulga et al., Abnormal bundling and accumulation of F-actin mediatestau-induced neuronal degeneration in vivo Nat Cell Biol. 9(2):139-48(2007)). In addition, fascin plays a role in Hodgkin's disease (Pinkuset al., Fascin, a sensitive new marker for Reed-Sternberg cells ofHodgkin's disease, Am J Pathol. 150(2):543-562 (1997)). Fascin alsoplays a role in processing and presenting antigens, for example, onantigen presenting cells (Mosialos et al., Circulating human dendriticcells differentially express high levels of a 55-kd actin-bundlingprotein. Am. J. Pathol. 148(2):593-600 (1996); Said et al. The role offollicular and inter digitating dendritic cells in HIV-related lymphoidhyperplasia: localization of fascin. Mod Pathol. 10(5):421-27 (1997)).Moreover, fascin also plays a role in ischemic injury (Meller et al.,Ubiquitin proteasome-mediated synaptic reorganization: a novel mechanismunderlying rapid ischemic tolerance, J Neurosci. 28(1):50-9 (2008)).

Provided herein are agents that modulate fascin activity and that can beused for methods of treating and inhibiting metastatic cancer, neuronaldisorders, neuronal degeneration, inflammatory conditions, viralinfections, bacterial infections, lymphoid hyperplasia, Hodgkin'sdisease, and ischemia-related tissue damage.

Tumor metastasis is the major cause of death of cancer patients (Weiss2000, Fidler 2003). Thus, inhibition or prevention of tumor metastasiswill significantly increase the survival rate of cancer patients, allowmore moderate radiation or chemotherapy with less side-effects, andcontrol the progression of solid tumors.

Tumor cell migration and invasion are critical steps in the process oftumor metastasis (Partin et al. 1989, Aznavoorian et a. 1993, Condeeliset al. 2005). For cell migration to proceed, the actin cytoskeleton mustbe reorganized by forming polymers and bundles to affect the dynamicchanges of cell shapes (Jaffe et al. 2005, Matsudaira 1994, Otto 1994).Individual actin filaments are flexible and elongation of individualfilaments per se is insufficient for membrane protrusion which isnecessary for cell migration. Bundling of actin filaments providesrigidity to actin filaments for protrusion against the compressive forcefrom the plasma membrane (Mogilner et al. 2005).

One of the critical actin-bundling proteins is fascin. Fascin is theprimary actin cross-linker in filopodia, which are membrane protrusionscritical for the migration and metastasis of cancer cells. Fascin isrequired to maximally cross-link the actin filaments into straight,compact, and rigid bundles. Elevated expressions of fascin mRNA andprotein in cancer cells have been correlated with aggressive clinicalcourse, poor prognosis and shorter survival. Accordingly, metastaticcancer can be treated, prevented and/or inhibited by administeringfascin inhibitors as described herein.

In addition, a cancer at any stage of progression can be treated by themethod of the present technology, such as primary, metastatic, andrecurrent cancers. In some embodiments, cancers are treated beforemetastasis is detected, for example, to inhibit metastatic cancer fromdeveloping. In other embodiments, cancers are treated when metastasis isdetected, for example, to inhibit further metastasis and progression ofthe cancer.

Compounds described herein, or pharmaceutically acceptable saltsthereof, can also be used to treat autoimmune deficiencysyndrome-associated Kaposi's sarcoma, cancer of the adrenal cortex,cancer of the cervix, cancer of the endometrium, cancer of theesophagus, cancer of the head and neck, cancer of the liver, cancer ofthe pancreas, cancer of the prostate, cancer of the thymus, carcinoidtumors, chronic lymphocytic leukemia, Ewing's sarcoma, gestationaltrophoblastic tumors, hepatoblastoma, multiple myeloma, non-small celllung cancer, retinoblastoma, or tumors in the ovaries. A cancer at anystage of progression can be treated or detected, such as primary,metastatic, and recurrent cancers. Information regarding numerous typesof cancer can be found, e.g., from the American Cancer Society(www.cancer.org), or from, e.g., Wilson et al. (1991) Harrison'sPrinciples of Internal Medicine, 12th Edition, McGraw-Hill, Inc.

In some embodiments, method are provided for treating or inhibitingmetastatic cancer in an animal, for example, for human and veterinaryuses, which include administering to a subject animal (e.g., a human), atherapeutically effective amount of a compound described herein, orpharmaceutically acceptable salt thereof. In some embodiments, the cellhas been removed from an animal.

Treatment of, or treating, a disease or condition (e.g., cancer) isintended to include the alleviation of or diminishment of at least onesymptom typically associated with the disease or condition. Thetreatment also includes alleviation or diminishment of more than onesymptom of the disease or condition. The treatment may cure the diseaseor condition, for example, by eliminating the symptoms and/or the sourceof the disease or condition. For example, treatment can cure the cancerby substantially inhibiting metastasis of the cancer cells so thatremoval or killing of the primary tumor or cancer cell(s) substantiallyeliminates the cancer. Treatment can also arrest or inhibit themetastasis of the cancer and/or tumor cells without directly killing orpromoting the apoptosis of cancer cells.

Fascin functions in a variety of cellular functions that play criticalroles in modulating the growth, movement and interaction of cells.However the actin bundling function of fascin is directly involved intumor metastasis and invasive growth.

The anti-metastatic activity of fascin (e.g., in the presence of varioustest agents or therapeutic agents like those described herein) can beevaluated against varieties of cancers using methods described hereinand available to one of skill in the art. Anti-cancer activity, forexample, can be determined by identifying the dose that inhibits 50%cancer cell metastasis (IC₅₀) of a compound or composition as describedherein.

Also provided is a method for evaluating a therapeutically effectivedosage for treating a cancer (e.g., inhibiting metastasis) with acompound described herein, or pharmaceutically acceptable salt thereof,that includes determining the IC₅₀ of the agent in vitro. Such a methodpermits calculation of the approximate amount of agent needed per volumeto inhibit cancer cell migration. Such amounts can be determined, forexample, by standard microdilution methods. In some embodiments, thecompound or composition as described herein can be administered inmultiple doses over an extended period of time, or intermittently.

Compositions

The compounds (e.g., fascin inhibitors) as described herein can beformulated as pharmaceutical compositions and administered to amammalian host, such as a human patient in a variety of forms adapted tothe chosen route of administration, i.e., orally or parenterally, byintravenous, intramuscular, topical, transdermally, intrathecally,ocularly, intranasally, intraperitoneally or subcutaneous routes.

The compounds (e.g., fascin inhibitors) described herein may besystemically administered, e.g., orally, in combination with apharmaceutically acceptable vehicle such as an inert diluent or anassimilable edible carrier. They may be enclosed in hard or soft shellgelatin capsules, may be compressed into tablets, or may be incorporateddirectly with the food of the patient's diet. For oral therapeuticadministration, the active compound may be combined with one or moreexcipients and used in the form of ingestible tablets, buccal tablets,troches, capsules, elixirs, suspensions, syrups, wafers, and the like.Such compositions and preparations should contain at least 0.1% ofactive compound. The percentage of the compositions and preparationsmay, of course, be varied and may conveniently be between about 2 toabout 60% of the weight of a given unit dosage form. The amount ofactive compound in such therapeutically useful compositions is such thatan effective dosage level will be obtained.

The tablets, troches, pills, capsules, and the like may also contain thefollowing: binders such as gum tragacanth, acacia, corn starch orgelatin; excipients such as dicalcium phosphate; a disintegrating agentsuch as corn starch, potato starch, alginic acid and the like; alubricant such as magnesium stearate; and a sweetening agent such assucrose, fructose, lactose or aspartame or a flavoring agent such aspeppermint, oil of wintergreen, or cherry flavoring may be added. Whenthe unit dosage form is a capsule, it may contain, in addition tomaterials of the above type, a liquid carrier, such as a vegetable oilor a polyethylene glycol. Various other materials may be present ascoatings or to otherwise modify the physical form of the solid unitdosage form. For instance, tablets, pills, or capsules may be coatedwith gelatin, wax, shellac or sugar and the like. A syrup or elixir maycontain the active compound, sucrose or fructose as a sweetening agent,methyl and propylparabens as preservatives, a dye and flavoring such ascherry or orange flavor. A material used in preparing any unit dosageform should be pharmaceutically acceptable and substantially non-toxicin the amounts employed. In addition, the active compound may beincorporated into sustained-release preparations and devices.

The active compounds described herein may also be administeredintravenously or intraperitoneally by infusion or injection. Solutionsof the active compound or its salts can be prepared in water, optionallymixed with a nontoxic surfactant. Dispersions can also be prepared inglycerol, liquid polyethylene glycols, triacetin, and mixtures thereofand in oils. Under ordinary conditions of storage and use, thesepreparations contain a preservative to prevent the growth ofmicroorganisms.

The pharmaceutical dosage forms suitable for injection or infusion caninclude sterile aqueous solutions or dispersions or sterile powderscomprising the active ingredient which are adapted for theextemporaneous preparation of sterile injectable or infusible solutionsor dispersions, optionally encapsulated in liposomes. In all cases, theultimate dosage form should be sterile, fluid and stable under theconditions of manufacture and storage. The liquid carrier or vehicle canbe a solvent or liquid dispersion medium comprising, for example, water,ethanol, a polyol (for example, glycerol, propylene glycol, liquidpolyethylene glycols, and the like), vegetable oils, nontoxic glycerylesters, and suitable mixtures thereof. The proper fluidity can bemaintained, for example, by the formation of liposomes, by themaintenance of the required particle size in the case of dispersions orby the use of surfactants. The prevention of the action ofmicroorganisms can be brought about by various antibacterial andantifungal agents, for example, parabens, chlorobutanol, phenol, sorbicacid, thimerosal, and the like. In many cases, it will be preferable toinclude isotonic agents, for example, sugars, buffers or sodiumchloride. Prolonged absorption of the injectable compositions can bebrought about by the use in the compositions of agents delayingabsorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions are prepared by incorporating the activecompound in the required amount in the appropriate solvent with severalof the other ingredients enumerated above, as required, followed byfilter sterilization. In the case of sterile powders for the preparationof sterile injectable solutions, the preferred methods of preparationare vacuum drying and the freeze drying techniques, which yield a powderof the active ingredient plus any additional desired ingredient presentin the previously sterile-filtered solutions.

For topical administration, the present compounds may be applied in pureform, i.e., when they are liquids. However, it will generally bedesirable to administer them to the skin as compositions orformulations, in combination with a dermatologically acceptable carrier,which may be a solid or a liquid.

Useful solid carriers include finely divided solids such as talc, clay,microcrystalline cellulose, silica, alumina and the like. Useful liquidcarriers include water, alcohols or glycols or water-alcohol/glycolblends, in which the present compounds can be dissolved or dispersed ateffective levels, optionally with the aid of non-toxic surfactants.Adjuvants such as fragrances and additional antimicrobial agents can beadded to optimize the properties for a given use. The resultant liquidcompositions can be applied from absorbent pads, used to impregnatebandages and other dressings, or sprayed onto the affected area usingpump-type or aerosol sprayers.

Thickeners such as synthetic polymers, fatty acids, fatty acid salts andesters, fatty alcohols, modified celluloses or modified mineralmaterials can also be employed with liquid carriers to form spreadablepastes, gels, ointments, soaps, and the like, for application directlyto the skin of the user.

Examples of useful dermatological compositions which can be used todeliver the compounds described herein, or pharmaceutically acceptablesalts thereof, to the skin are known to the art; for example, seeJacquet et al. (U.S. Pat. No. 4,608,392), Geria (U.S. Pat. No.4,992,478), Smith et al. (U.S. Pat. No. 4,559,157) and Wortzman (U.S.Pat. No. 4,820,508).

Useful dosages of the compounds described herein, or pharmaceuticallyacceptable salts thereof, can be determined by comparing their in vitroactivity, and in vivo activity in animal models. Methods for theextrapolation of effective dosages in mice, and other animals, to humansare known to the art; for example, see U.S. Pat. No. 4,938,949.

Generally, the concentration of the compounds described herein, orpharmaceutically acceptable salts thereof, in a liquid composition, suchas a lotion, will be about 0.01 wt %, about 0.1 wt %, about 1.0 wt %,about 2.0 wt %, about 3.0 wt %, about 4.0 wt %, about 5.0 wt %, about10.0 wt %, about 25.0 wt %, or a range between and including any two ofthese values. The concentration in a semi-solid or solid compositionsuch as a gel or a powder will be about 0.01 wt %, about 0.1 wt %, about1.0 wt %, about 2.0 wt %, about 3.0 wt %, about 4.0 wt %, about 5.0 wt%, about 10.0 wt %, about 25.0 wt %, or a range between and includingany two of these values.

The amount of the compound, or an active salt or derivative thereof,required for use in treatment will vary not only with the particularsalt selected but also with the route of administration, the nature ofthe condition being treated and the age and condition of the patient andwill be ultimately at the discretion of the attendant physician orclinician. In general, however, a suitable dose will be in the range offrom about 1.0 to about 200 mg/kg, e.g., from about 1 to about 100 mg/kgof body weight per day, such as about 2.0 to about 100 mg/kg of bodyweight per day, such as about 3.0 to about 50 mg per kilogram bodyweight of the recipient per day, or in the range of about 5 to 20mg/kg/day. Alternatively, the compositions can be administered fivetimes a week on five consecutive days with a two day rest, or four timesa week on four consecutive days with a three day rest, or every otherday.

Methods for extrapolating effective dosages in mice and other animals,to humans are known in the art (See, for example, U.S. Pat. No.4,938,949). For example, in some embodiments, compounds describedherein, or pharmaceutically acceptable salts thereof, (for example thoseuseful for the treatment of colon and/or ovarian cancer) may beadministered at dosage levels of about 0.01 mg/kg to about 300 mg/kg,from about 0.1 mg/kg to about 250 mg/kg, from about 1 mg/kg to about 200mg/kg, from about 1 mg/kg to about 150 mg/kg, from about 1 mg/kg toabout 100 mg/kg, from about 1 mg/kg to about 90 mg/kg, from about 1mg/kg to about 80 mg/kg, from about 1 mg/kg to about 70 mg/kg, fromabout 1 mg/kg to about 60 mg/kg, from about 1 mg/kg to about 50 mg/kg,from about 1 mg/kg to about 40 mg/kg, from about 1 mg/kg to about 30mg/kg, from about 1 mg/kg to about 20 mg/kg, from about 5 mg/kg to about100 mg/kg, from about 5 mg/kg to about 90 mg/kg, from about 5 mg/kg toabout 80 mg/kg, from about 5 mg/kg to about 70 mg/kg, from about 5 mg/kgto about 60 mg/kg, from about 5 mg/kg to about 50 mg/kg, from about 5mg/kg to about 40 mg/kg, from about 5 mg/kg to about 30 mg/kg, fromabout 5 mg/kg to about 20 mg/kg, from about 10 mg/kg to about 100 mg/kg,from about 10 mg/kg to about 90 mg/kg, from about 10 mg/kg to about 80mg/kg, from about 10 mg/kg to about 70 mg/kg, from about 10 mg/kg toabout 60 mg/kg, from about 10 mg/kg to about 50 mg/kg, from about 10mg/kg to about 40 mg/kg, from about 10 mg/kg to about 30 mg/kg, fromabout 10 mg/kg to about 20 mg/kg, from about 20 mg/kg to about 100mg/kg, from about 20 mg/kg to about 90 mg/kg, from about 20 mg/kg toabout 80 mg/kg, from about 20 mg/kg to about 70 mg/kg, from about 20mg/kg to about 60 mg/kg, from about 20 mg/kg to about 50 mg/kg, fromabout 20 mg/kg to about 40 mg/kg, from about 20 mg/kg to about 30 mg/kg,of subject body weight per day, one or more times a day, to obtain thedesired therapeutic effect. In some embodiments, compounds may beadministered at a dosage of about 1 mg/kg or greater, 5 mg/kg orgreater; 10 mg/kg or greater, 15 mg/kg or greater, 20 mg/kg or greater,25 mg/kg or greater, 30 mg/kg or greater, 35 mg/kg or greater, 40 mg/kgor greater, 45 mg/kg or greater, 50 mg/kg or greater, 60 mg/kg orgreater, 70 mg/kg or greater, of body weight. It will also beappreciated that dosages smaller than 0.01 mg/kg or greater than 70mg/kg (for example 70-200 mg/kg) can be administered to a subject.

In some embodiments, the compounds described herein may be used inchemotherapy (i.e., to inhibit metastasis) and may be administered athigher dosage. For example, compounds to be used in chemotherapy may beadministered from about 100 mg/kg to about 300 mg/kg, from about 120mg/kg to about 280 mg/kg, from about 140 mg/kg to about 260 mg/kg, fromabout 150 mg/kg to about 250 mg/kg, from about 160 mg/kg to about 240mg/kg, of subject body weight per day, one or more times a day, toobtain the desired therapeutic effect.

In certain other embodiments, the compounds described herein may be usedin supportive therapy (e.g., as an adjuvant to surgery or irradiation ina range of common types of tumor) and may be administered at lowerdosage. For example, compounds to be used in supportive therapy may beadministered from about 1 mg/kg to about 30 mg/kg, from about 1 mg/kg toabout 25 mg/kg, from about 5 mg/kg to about 20 mg/kg, of subject bodyweight per day, one or more times a day, to obtain the desiredtherapeutic effect.

In certain other embodiments, the compounds described herein may be usedfor treating metastatic cancer (e.g., ovarian and/or colon cancer) andmay be administered at an intermediate dosage. For example, compounds tobe used in supportive therapy may be administered from about 1 mg/kg toabout 100 mg/kg, from about 1 mg/kg to about 80 mg/kg, from about 5mg/kg to about 70 mg/kg, from about 10 mg/kg to about 70 mg/kg, fromabout 10 mg/kg to about 60 mg/kg, from about 20 mg/kg to about 70 mg/kg,from about 20 mg/kg to about 60 mg/kg, of subject body weight per day,one or more times a day, to obtain the desired therapeutic effect.

The compound is conveniently administered in unit dosage form; forexample, containing 45 to 3000 mg, conveniently 90 to 2250 mg, mostconveniently, 450 to 1500 mg of active ingredient per unit dosage form.In some embodiments, the compound is administered at dosages of about 1to about 100 mg/kg.

Ideally, the active ingredient should be administered to achieve peakplasma concentrations of the active compound of from about 0.5 nM toabout 10 μM, or about 1 nM to 1 μM, or about 10 nM to about 0.5 μM. Thismay be achieved, for example, by the intravenous injection of a 0.05 to5% solution of the active ingredient, optionally in saline, or orallyadministered as a bolus containing about 20-2000 mg of the activeingredient. Desirable blood levels may be maintained by continuousinfusion to provide about 0.2 to 1.0 mg/kg/hr or by intermittentinfusions containing about 0.4 to 20 mg/kg of the active ingredient(s).The desired dose may conveniently be presented in a single dose or asdivided doses administered at appropriate intervals, for example, astwo, three, four or more sub-doses per day. The sub-dose itself may befurther divided, e.g., into a number of discrete loosely spacedadministrations; such as multiple inhalations from an insufflator or byapplication of a plurality of drops into the eye.

Compounds described herein, or pharmaceutically acceptable saltsthereof, are useful as therapeutic agents administered for inhibition ofcell migration and treatment of metastatic cancer. Such cancers includebut are not limited to, e.g., cancers involving the animal's head, neck,lung, mesothelioma, mediastinum, esophagus, stomach, pancreas,hepatobiliary system, small intestine, colon, colorectal, rectum, anus,kidney, ureter, bladder, prostate, urethra, penis, testis, gynecologicalorgans, ovaries, breast, endocrine system, skin, or central nervoussystem. Thus, for example, the cancer can be a breast cancer, aleukemia, a lung cancer, a colon cancer, a central nervous systemcancer, a melanoma, an ovarian cancer, a renal cancer, or a prostatecancer.

Additionally, compounds described herein, or pharmaceutically acceptablesalts thereof, such as the exemplary salts described herein, may beuseful as pharmacological tools for the further investigation of theinhibition of cell migration.

The compounds described herein, or pharmaceutically acceptable saltsthereof, can also be administered in combination with other therapeuticagents that are effective for treating or controlling the spread ofcancerous cells or tumor cells.

Moreover, the compounds described herein, or pharmaceutically acceptablesalts thereof, can be tested in appropriate animal models. For example,the compounds described herein, or pharmaceutically acceptable saltsthereof, can be tested in animals with known tumors, or animals thathave been injected with tumor cells into a localized area. The degree ornumber of secondary tumors that form over time is a measure ofmetastasis and the ability of the compounds to inhibit such metastasiscan be evaluated relative to control animals that have the primary tumorbut receive no test compounds.

The compounds described herein, or pharmaceutically acceptable saltsthereof, will also find use in treatment of brain disorders (Kraft etal., J. Neurosci. 2006 Aug. 23; 26(34):8734-47); Hodgkin's disease(Pinkus et al., Am J Pathol. 1997 February; 150(2):543-62); virusinfection (Mosialos et al., Am J Pathol. 1996 February; 148(2):593-600);neuronal degeneration (Fulga et al., Nat Cell Biol. 2007 February: 9(2):139-48); lymphoid hyperplasia (Said et al., Mod Pathol. 1997 May;10(5):421-7); and ischemia (Meller et al., J Neurosci. 2008 Jan. 2;28(1):50-9.)

General Synthetic Methods

The compounds described herein are commercially available or can beprepared from readily available starting materials using the followinggeneral methods and procedures. It will be appreciated that wheretypical or preferred process conditions (i.e., reaction temperatures,times, mole ratios of reactants, solvents, pressures, etc) are given,other process conditions can also be used unless otherwise stated.Optimum reaction conditions may vary with the particular reactants orsolvent used, but such conditions can be determined by one skilled inthe art by routine optimization procedures.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary to prevent certainfunctional groups from undergoing undesired reactions. Suitableprotecting groups for various functional groups as well as suitableconditions for protecting and deprotecting particular functional groupsare well known in the art. For example, numerous protecting groups aredescribed in T. W. Greene and G. M. Wuts, Protecting Groups in OrganicSynthesis, Third Edition, Wiley, New York, 1999, and references citedtherein.

Furthermore, the compounds described herein may contain one or morechiral centers. Accordingly, if desired, such compounds can be preparedor isolated as pure stereoisomers, i.e., as individual enantiomers ordiastereomers, or as stereoisomer-enriched mixtures. All suchstereoisomers (and enriched mixtures) are included within the scope ofthis invention, unless otherwise indicated. Pure stereoisomers (orenriched mixtures) may be prepared using, for example, optically activestarting materials or stereoselective reagents well-known in the art.Alternatively, racemic mixtures of such compounds can be separatedusing, for example, chiral column chromatography, chiral resolvingagents and the like.

The starting materials for the following reactions are generally knowncompounds or can be prepared by known procedures or obviousmodifications thereof. For example, many of the starting materials areavailable from commercial suppliers such as Aldrich Chemical Co.(Milwaukee, Wis., USA), Bachem (Torrance, Calif., USA), Emka-Chemce orSigma (St. Louis, Mo., USA). Others may be prepared by procedures, orobvious modifications thereof, described in standard reference textssuch as Fieser and Fieser's Reagents for Organic Synthesis, Volumes 1-15(John Wiley and Sons, 1991), Rodd's Chemistry of Carbon Compounds,Volumes 1-5 and Supplementals (Elsevier Science Publishers, 1989),Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991), March'sAdvanced Organic Chemistry, (John Wiley and Sons, 4^(th) Edition), andLarock's Comprehensive Organic Transformations (VCH Publishers Inc.,1989).

The various starting materials, intermediates, and compounds describedherein may be isolated and purified where appropriate using conventionaltechniques such as precipitation, filtration, crystallization,evaporation, distillation, and chromatography. Characterization of thesecompounds may be performed using conventional methods such as by meltingpoint, mass spectrum, nuclear magnetic resonance, and various otherspectroscopic analyses.

Schemes 1-5 show exemplifying methods for preparing compounds describedherein.

In Scheme 1, Compound 1-1, wherein Lg is a leaving group, such asfluoro, chloro, bromo, iodo, tosylate, triflate, and the like, reactswith R¹—OH, R¹—SH or R¹—HNR⁸ to form Compound 1-2 wherein L¹ is —O—, —S—or —NR⁸—, respectively. Alternatively, Lg is hydrogen and Y³ is nitrogenin Compound 1-1, which reacts with R¹-L¹-Lg², wherein Lg² is a leavinggroup, such as halo, to form Compound 1-2, wherein L¹ is as defined inFormula I-a, such as —C(R⁸)₂—.

Compound 1-2 is reduced by a reducing agent, such as H₂ in the presenceof a catalyst (e.g., Pd) to Compound 1-3. In some embodiments, Compound1-3 reacts with a carboxylic acid compound R⁵CO₂H under couplingconditions, such as using an amide coupling reagent, to form a compoundof Formula I-a wherein R is L²-R⁵ and L² is —NHC(O)—. In someembodiments, Compound 1-3 can react with a sulfonylchloride compoundR⁵SO₂Cl to form a compound of Formula I-a wherein R is L²-R⁵ and L² is—NHS(O)₂—. In some embodiments, Compound 1-3 reacts with sodium nitriteand a nucleophile Z⁻M⁺, wherein Z⁻ is a nucleophile such as iodide,bromide, chloride, fluoride, cyanide, carboxyl, and M⁺ is the counterion, such as copper, sodium, etc., to form various intermediatesrepresented by Compound 1-4. Compound 1-4 can undergo a variety oftransformations to provide for embodiments of the compound of FormulaI-a. For example, when Z is iodo, bromo or chloro, Compound 1-4 canundergo various cross-coupling reactions with another iodo, bromo orchloro compound, or a compound having a boronic acid functionality toform a compound of Formula I-a wherein R is L²-R⁵ and L² is a covalentbond. Preferably, such reactions are conducted under catalyticconditions using a catalyst such as CuI or a palladium catalyst.Alternatively, when Z is carboxyl, Compound 1-4 can react with an amineR⁵—NHR⁸ to form a compound of Formula I-a wherein R is L²-R⁵ and L² is—C(O)N(R⁸)—. Still alternatively, Compound 1-3 reacts with sodiumnitrite and sulfur dioxide and copper (I) chloride to form asulfonylchloride compound (Z is SO₂Cl) which can react with an amineR⁵—NHR⁸ to form a compound of Formula I-a wherein R is L²-R⁵ and L² is—S(O)₂N(R⁸)—. In Scheme 1, Q¹, Q², R¹, R⁴, R⁵, X¹, Y¹, Y², Y³, Y⁵, Y⁶, sand t are as defined in Formula I-a.

Amide coupling reagents are known in the art and may include, but arenot limited to, amininum and phosphonium based reagents. Aminium saltsincludeN-[(dimethylamino)-1H-1,2,3-triazolo[4,5-b]pyridine-1-ylmethylene]-N-methylmethanaminiumhexafluorophosphate N-oxide (HATU),N-[(1H-benzotriazol-1-yl)(dimethylamino)methylene]-N-methylmethanaminiumhexafluorophosphate N-oxide (HBTU),N-[(1H-6-chlorobenzotriazol-1-yl)(dimethylamino)methylene]-N-methylmethanaminiumhexafluorophosphate N-oxide (HCTU),N-[(1H-benzotriazol-1-yl)(dimethylamino)methylene]-N-methylmethanaminiumtetrafluoroborate N-oxide (TBTU), andN-[(1H-6-chlorobenzotriazol-1-yl)(dimethylamino)methylene]-N-methylmethanaminiumtetrafluoroborate N-oxide (TCTU).

Phosphonium salts include7-azabenzotriazol-1-yl-N-oxy-tris(pyrrolidino)phosphoniumhexafluorophosphate (PyAOP) andbenzotriazol-1-yl-N-oxy-tris(pyrrolidino)phosphonium hexafluorophosphate(PyBOP). Amide formation step may be conducted in a polar solvent suchas dimethylformamide (DMF) and may also include an organic base such asdiisopropylethylamine (DIEA) or dimethylaminopyridine (DMAP).

Cross-coupling reactions are well known in the art and, for example, arereported in Anna Roglans, et al. Diazonium Salts as Substrates inPalladium-Catalyzed Cross-Coupling Reactions, Chem. Rev., 2006, 106(11):4622-4643; Brad M. Rosen, et al., Nickel-Catalyzed Cross-CouplingsInvolving Carbon-Oxygen Bonds, Percec Chem. Rev., 2011, 111(3):1346-1416; Jean-Pierre Corbet, et al., Selected PatentedCross-Coupling Reaction Technologies, Chem. Rev., 2006, 106(7):2651-2710; Gwilherm Evano et al., Copper-Mediated Coupling Reactionsand Their Applications in Natural Products and Designed BiomoleculesSynthesis, Chem. Rev., 2008, 108 (8):3054-3131; Benny Bogoslavsky, etal., Formation of a Carbon-Carbon Triple Bond by Coupling Reactions InAqueous Solution, Science 308 (5719): 234-235 (2005); and M. Lafrance,et al., Catalytic Intermolecular Direct Arylation of Perfluorobenzenes,J. Am. Chem. Soc. 128 (27): 8754-8756 (2006); Norio Miyaura, et al., “Anew stereospecific cross-coupling by the palladium-catalyzed reaction of1-alkenylboranes with 1-alkenyl or 1-alkynyl halides,” TetrahedronLetters, 1979, 20(36): 3437-3440; P. E. Fanta, “The Ullmann Synthesis ofBiaryls”, Synthesis, 1974, 1974: 9-21; M. Gomberg, and W. E. Bachmann,J. Am. Chem. Soc., 1924, 42(10):2339-2343; R. J. P. Corriu and Masse, J.P. “Activation of Grignard reagents by transition-metal complexes. A newand simple synthesis of trans-stilbenes and polyphenyls,” Journal of theChemical Society, Chemical Communications, 1972, (3):144a.

In Scheme 2, Compound 2-1, wherein Lg is a leaving group, such asfluoro, chloro, bromo, iodo, tosylate, triflate, and the like, andwherein R^(P) is hydrogen or a carboxy protecting group such as loweralkyl or benzyl, reacts with R¹—OH, R¹—SH or R¹—HNR⁸ to form theCompound 2-2. When R^(p) a carboxy protecting group, Compound 2-2 can bedeprotected to Compound 2-3. Compound 2-2 or Compound 2-3 can react withoptionally substituted aniline under coupling conditions, such as usingan amide coupling reagent, to form a compound of Formula I-c wherein L²is —C(O)NR⁸—. In Scheme 2, R¹, R², R³, L¹, X¹, m and n are as defined inFormula I-c.

In Scheme 3, Compound 3-1, wherein Lg is a leaving group, such asfluoro, chloro, bromo, iodo, tosylate, triflate, and the like, reactswith R¹—OH, R¹—SH or R¹—HNR⁸ to form Compound 3-2, which is reduced by areducing agent, such as H₂ in the presence of a catalyst (e.g., Pd) toCompound 3-3. Compound 3-3 then reacts with an optionally substitutedbenzoic acid under coupling conditions, such as using an amide couplingreagent, to form a compound of Formula I-c wherein L² is —NHC(O)—.Alternatively, Compound 3-3 can react with an optionally substitutedbenzenesulfonylchloride to form a compound of Formula I-c wherein L² is—NHS(O)₂—. In Scheme 3, R¹, R², R³, L¹, X¹, m and n are as defined inFormula I-c.

A compound of Formula I-c can have a tautomer having a structure ofFormula I-d, wherein L³ is ═NC(O)— or ═NSO₂— and X² is O, S or NR⁸.

In Scheme 4, Compound 4-1, Pr is an amino protecting group, such astert-butoxycarbonyl or carboxybenzyl, Z¹ and Z² are groups suitable fora cross-coupling reaction, such as iodo, bromo, chloro, tosylate,boronic acid, etc. The other variables are as defined in Formula II. Z¹and Z² can be the same or different. When Z¹ and Z² are the same,Compound 4-2 wherein R²¹ and R²² are the same can be prepared through aone-step cross-coupling reaction by reacting with at least twoequivalents of a compound of the formula R²¹Z³, wherein Z³ is a groupthat can react with Z¹ and Z² in the cross-coupling reaction.Alternatively, Z¹ and Z² are different and Compound 4-2 wherein R²¹ andR²² are different can be prepared through a first cross-couplingreaction using conditions that can selectively couple the first of R²¹and R²² followed by a second cross-coupling reaction to couple thesecond of R²¹ and R²². The amino protecting group of Compound 4-2 can bedeprotected under conditions known in the art to provide for a compoundof Formula II wherein

is a single bond and R²⁴ is hydrogen, which may exist as its tautomerwherein

is a double bond and R²⁴ is absent, or may be alkylated with a lowerhaloalkyl to form a compound of Formula II wherein

is a single bond and R²⁴ is lower alkyl.

In Scheme 5, Compound 5-1 reacts with R³⁰—CO₂H under couplingconditions, such as using an amide coupling reagent, to form a compoundof Formula III wherein X³⁰ is C(═O). Alternatively, Compound 5-1 reactswith R³⁰—COCl or R³⁰—SO₂Cl under basic conditions, such as using anorganic base, e.g., triethylamine, diisopropylethylamine or pyridine, toform a compound of Formula III wherein X³⁰ is C(═O) or SO₂. Thevariables in Scheme 5 are as defined in Formula III.

All publications, patent applications, issued patents, and otherdocuments referred to in this specification are herein incorporated byreference as if each individual publication, patent application, issuedpatent, or other document was specifically and individually indicated tobe incorporated by reference in its entirety. Definitions that arecontained in text incorporated by reference are excluded to the extentthat they contradict definitions in this disclosure.

The present technology, thus generally described, will be understoodmore readily by reference to the following Examples, which is providedby way of illustration and is not intended to be limiting of the presenttechnology.

EXAMPLES Example 1: High Throughput Fascin Inhibition Assay

A high throughput assay was developed to screen for fascin specificinhibitors. Purified polymerized F-actin with or without fascin weremixed and incubated to allow actin bundle formation. F-actin polymerswere then bound to the poly-D-lysine coated plates. After extensivewashes, F-actin polymers were visualized by labeling them with AlexaFluor 488 phalloidin. Four images were taken from each well and theaverage fiber length was analyzed for each tested compound. In thepresence of fascin, actin fibers were longer and thicker as shown below.30 μM of2-chloro-N-(6-chlorobenzo[d]thiazol-2-yl)-5-nitrobenzenesulfonamide(Compound 3) inhibited fiber growth.

Average fiber length Average fiber thickness (arbitrary unit) (arbitraryunit) Actin 0.36 0.32 Actin + Fascin 1.57 0.53 Actin + Fascin + 0.390.35 Compound 3 (30 μM)

From the primary screens of ˜150,000 small molecule compounds, ˜700compounds were identified that resulted in shorter and thinner actinbundles when compared with those without the small molecule compounds.

The images from screens with these ˜700 compounds were individuallyexamined to insure that they looked like those of actin alone controls.After 2^(nd) and 3^(rd) confirmative screens, in which both muscle andnon-muscle actin proteins were tested, 145 small-molecule compounds wereconfirmed to inhibit the actin-bundling function of fascin. Thedose-response curves of these 145 compounds were established.

Example 2: Human Fascin-1 Expression and Purification

Recombinant human fascin 1 was expressed as a GST fusion protein in BL21Escherichia coli. One liter of 2YT medium with ampicillin was inoculatedovernight with 3 mL of BL21/DE3 culture transformed with pGEX4T-fascin 1plasmid and grown at 37° C. until attenuance at 600 nm (D600) reachedabout 0.8. The culture was then transferred to 18° C. and induced by theaddition of 0.1 mM isopropyl 3-d-thiogalactoside (IPTG) for 12 h.Bacteria were harvested by centrifugation at 5,000 r.p.m. for 10 min.The pellets were suspended in 30 mL of PBS supplemented with 0.2 mMPMSF, 1 mM DTT, 1% (v/v) Triton X-100 and 1 mM EDTA. After sonication,the suspension was centrifuged at 15,000 r.p.m. for 30 min to remove thecell debris. The supernatant was then incubated for 2 h with 4 mL ofglutathione beads (Sigma) at 4° C. After extensive washing with PBS, thebeads were resuspended in 10 mL of thrombin cleavage buffer (20 mMTris-HCl pH 8.0, 150 mM NaCl, 2 mM CaCl₂, 1 mM DTT). Fascin was releasedfrom the beads by incubation overnight with 40-100 U of thrombin at 4°C. After centrifugation, 0.2 mM PMSF was added to the supernatant toinactivate the remnant thrombin activity. The fascin protein was furtherconcentrated with a Centricon® (Boca Raton, Fla.) filter to about 50mg/mL.

Example 3: Quantification of Fascin Expression Levels

The levels of fascin mRNA and protein can be determined by real-time PCRand Western blot, respectively. For quantitative real-time PCR, samplesfrom cancer patients were used for RNA isolation. Oligonucleotideprimers specific for fascin mRNA were used for PCR reactions. ForWestern blots, samples from cancer patients were assessed withanti-fascin antibody. The intensity of the bands representing fascinproteins was quantified by image documentation and quantificationsoftware.

Example 4: Chemical Library Screens and Analysis

About 150,000 compounds were screened. These chemical compounds werefrom the LOPAC 1280 collection, the Prestwick chemical library, thePharmakon collection, the MicroSource Spectrum collection, the LifeChemicals library, the Greenpharma natural compound library, the Enaminelibrary, the ChemBridge library, the Chem-X-Infinity library, and theBioFocus DPI library. Purified fascin protein (15 μL of 0.5 μM) inbuffer (100 mM KCl, 20 mM Tris/HCl, pH 7.5, 2 mM MgCl₂) was added intoeach well of a clear 384-well flat-bottom plate (Corning) using ThermoMultidrop Combi (Fisher). Compound (180 nL) solutions (5 mM stock) fromvarious chemical libraries were pin transferred from stock 384-wellplates into the 384-well assay plates and incubated for 30 min. Then 15μL of 0.5 μM polymerized actin (in 100 mM KCl, 20 mM Tris/HCl, pH 7.5, 2mM MgCl₂, 1 mM DTT, 1 mM ATP) (Cytoskeleton Inc.) was added, resultingin 30 μM final concentration for chemical compounds. After another 30min, 10 μL of Alexa Fluro 488 Phalloidin (25 times dilution from stocksin 100% methanol, Invitrogen) was added to stain F-actin and wasincubated in the dark for one hour. Mixed solution (25 μL) was thentransferred to one well in a black 384-well plate coated withpoly-D-lysine, and stained actin bundles or F-actin would stick onto thepoly-D-lysine plates. After the plates were thoroughly washed with 1×PBSfor 3 times, the plate was imaged using an ImageXpress Micro HighContent Screening System (Molecular devices). The images were processedand analyzed using MetaMorph software. The raw image data for each wellwas background-corrected by subtraction of the median intensities acrossall wells on the plate. The background-corrected data was used tocompute the bundle length for each well. The negative control wells wereemployed for quality control: multiple DMSO-only control wells (16wells/plate) were present on each assay plate. The top ten compoundswith the shortest bundle length on each plate were chosen for subsequentconfirmative screens. In the confirmative screens, ˜700 compounds weretested in duplicate. One hundred and forty-five compounds with confirmedresponses were picked and preceded to the IC₅₀ studies.

In confirmative screening of selected compounds, a control with anotheractin-bundling protein, fimbrin, was used to eliminate compounds thatare not specific to fascin. Also in confirmative screening, eachcompound was tested in duplicate on the same plate.

The concentration of the test compounds varied in certain assays.

Certain compounds of Formula I-a, I-b, II or III and their IC₅₀ areshown below in Table 1.

TABLE 1 Inhibition of Fascin Activity Inhibition of Fascin Activity #Name Structure IC₅₀ (μM)  1 5-(3,4-dichlorobenzyl)-1- (S,S-dioxo-tetrahydrothiophen-3-yl)- 1H-pyrazolo[3,4- d]pyrimidin-4(5H)-one

 0.734  2 N-(1-(4- (trifluoromethyl)benzyl)- 1H-indazol-3-yl)furan-2-carboxamide

2   3 2-chloro-N-(6- chlorobenzo[d]thiazol-2- yl)-5-nitrobenzenesulfonamide

 0.652  4 (Z)-N-(2,3-diphenyl-1,2,4- thiadiazol-5(2H)-ylidene)methanamine

0.5  5 N-methyl-2,3-diphenyl- 1,2,4-thiadiazolium-5- amine

0.5  6 N-benzyl-2,3-diphenyl- 1,2,4-thiadiazolium-5- amine

5   7 N-phenyl-2,3-diphenyl- 1,2,4-thiadiazolium-5- amine

2   8 N-(3-(1H-1,2,4-triazol-3- ylthio)-4- hydroxynaphthalen-1-yl)- 2,5-dimethylbenzenesulfonamide

 0.683  9 N-(3-(1H-(1,2,4-triazol-3- ylthio)-4- hydroxynaphthalen-1-yl)-4- ethoxybenzenesulfonamide

 1.47 10 N-(3-(1H-1,2,4-triazol-3- ylthio)-4- hydroxynaphthalen-1-yl)-4- methoxybenzenesulfonamide

 0.807 11 N-(3-(1H-1,2,4-triazol-3- ylthio)-4- hydroxynaphthalen-1-yl)-4- ethylbenzenesulfonamide

 0.89 12 N-(3-(1H-1,2,4-triazol-3- ylthio)-4- hydroxynaphthalen-1-yl)-2,4,5- trimethylbenzenesulfonamide

 0.539 13 (Z)-N-(3-(1H-1,2,4-triazol- 3-ylthio)-4-oxonaphthalen- 1(4H)-ylidene)benzenesulfonamide

2  14 N-(3-(1H-1,2,4-triazol-3- ylthio)-4- hydroxynaphthalen-1-yl)- 4-bromobenzenesulfonamide

 2.62 15 N-(3-(1H-1,2,4-triazol-3- ylthio)-4- hydroxynaphthalen-1-yl)-2,4- dimethylbenzenesulfonamide

2.7 16 3-chloro-N- (6-phenoxybenzo[d]thiazol-2- yl)benzenesulfonamide

about 4  17 N-(6- fluorobenzo[d]thiazol-2- yl)-3-nitrobenzenesulfonamide

about 4  18 2,3-dichloro-N-(6- fluorobenzo[d]thiazol-2-yl)benzenesulfonamide

about 13 19 N-(6- chlorobenzo[d]thiazol-2- yl)acetamide

about 2  20 N-(benzo[d]thiazol-2- yl)cinnamamide

about 4  21 5-(3-chlorobenzyl)-1-(2- hydroxyethyl)-1H-pyrazolo[4,3-d]pyrimidin- 4(5H)-one

1.2 23 2-(4-oxo-1-(S,S-dioxo- tetrahydrothiophen-3-yl)- 1H-pyrazolo[3,4-d]pyrimidin-5(4H)-yl)-N- (3- (trifluoromethyl)phenyl) acetamide

3.4 24 N-(4-fluorobenzyl)-2-(4- oxo-1-(S,S-dioxo-tetrahydrothiophen-3-yl)- 1H-pyrazolo[3,4- d]pyrimidin-5(4H)-yl)acetamide

4.3 25 N-(benzo[d][1,3]dioxol-5- ylmethyl)-2-(4-oxo-1-(S,S-dioxo-tetrahydrothiophen- 3-yl)-1H-pyrazolo[3,4- d]pyrimidin-5(4H)-yl)acetamide

4.6 26 N-(4-chlorophenyl)-2-(4- oxo-1-(S,S-dioxo-tetrahydrothiophen-3-yl)- 1H-pyrazolo[3,4- d]pyrimidin-5(4H)-yl)acetamide

4.3 27 5-(2-(2,3- dihydrobenzo[b][1,4]dioxin- 6-yl)-2-oxoethyl)-1-(S,S-dioxo-tetrahydrothiophen- 3-yl)-1H-pyrazolo[3,4- d]pyrimidin-4(5H)-one

about 10 28 5-(2-(2,4-dimethylphenyl)- 2-oxoethyl)-1-(S,S-dioxo-tetrahydrothiophen-3-yl)- 1H-pyrazolo[3,4- d]pyrimidin-4(5H)-one

4.7 29 5-(2-(benzo[d][1,3]dioxol- 5-yl)-2-oxoethyl)-1-(S,S-dioxo-tetrahydrothiophen- 3-yl)-1H-pyrazolo[3,4- d]pyrimidin-4(5H)-one

3.7 30 5-(3,4-dichlorobenzyl)-1- o-tolyl-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one

2.0 31 5-(3,4-dichlorobenzyl)-1- (2,3-dimethylphenyl)-1H-pyrazolo[3,4-d]pyrimidin- 4(5H)-one

2.3 32 5-(3,4-dichlorobenzyl)-1- (2,4-dimethylphenyl)-1H-pyrazolo[3,4-d]pyrimidin- 4(5H)-one

2.1

Based on their in vitro fascin inhibitory activities, the abovecompounds are contemplated to be used for treating a condition ordisorder mediated by fascin activity.

Example 5: Boyden-Chamber Cell Migration Assay

Since fascin is critical for tumor cell migration, inhibitors of fascinshould block tumor cell migration. Representative compounds wereselected from the above-identified fascin inhibitors to test theirabilities on inhibition of tumor cell migration (See Table 2). Boydenchamber assays for cell migration were used to show that these compoundsinhibited the migration of breast tumor cells, prostate tumor cells, andlung tumor cells. Therefore, tumor cells with fascin expression arelikely sensitive to these fascin inhibitors. The cell lines used arelisted below.

4T1 breast tumor cells MDA-MB-231 breast tumor cells DU145 prostatetumor cells PC-3 prostate tumor cells LLC lung tumor cells

Exemplifying procedure: MDA-MB-231 cells (5×10⁴) or 4T1 Cells (1×10⁵)suspended in 100 μL starvation medium were added to the upper chamber ofan insert (6.5 mm diameter, 8 m pore size; Becton Dickson), and theinsert was placed in a 24-well plate containing 700 μL starvation mediumwith or without 10% FBS. When used, inhibitors were added to the lowerchamber. Migration assays were performed for 6 h and cells were fixedwith 3.7% formaldehyde. Cells were stained with crystal violet stainingsolution, and cells on the upper side of the insert were removed with acotton swab. Three randomly selected fields (×10 objectives) on thelower side of the insert were photographed, and the migrated cells werecounted. Migration was expressed as average number of migrated cells ina field.

TABLE 2 Inhibition of Fascin Activity and Tumor Cell Migration CompoundStructure Assay IC₅₀ (μM)*  3

4T1 breast tumor cells MDA-MB-231 breast tumor cells DU145 prostatetumor cells PC-3 prostate tumor cells LLC lung tumor cells 20 25   25 2525  4

4T1 breast tumor cells MDA-MB-231 breast tumor cells 50 30  5

4T1 breast tumor cells 30  7

4T1 breast tumor cells >50   8

4T1 breast tumor cells 20  9

4T1 breast tumor cells >20  10

4T1 breast tumor cells MDA-MB-231 breast tumor cells 19 20 11

4T1 breast tumor cells ~20  12

4T1 breast tumor cells 16 13

4T1 breast tumor cells >30  14

4T1 breast tumor cells ~25  15

4T1 breast tumor cells >30  16

4T1 breast tumor cell about 7  17

4T1 breast tumor cell about 60 18

4T1 breast tumor cell about 3  19

4T1 breast tumor cell about 25 20

4T1 breast tumor cell about 60 Statistical Analysis: Data are expressedas mean ± S.D. and analyzed by Student's t test with significancedefined as p < 0.05.

The compounds shown above were representative of those that inhibitedtumor cell migration. In vitro data obtained in such assays are known tocorrelate with results obtained from in vivo models. See, e.g., Shan,D., et al., Synthetic analogues of migrastatin that inhibit mammarytumor metastasis in mice, Proc. Nat. Acad. Sci. 102: 3772-3776 (2005).Based on their in vitro cell migration inhibition activities and knowncorrelation with in vivo activities, the compounds are contemplated tobe useful for treating a condition or disorder mediated by fascinactivity and/or tumor metastasis.

Example 6: Tumor Metastasis in Mouse Models

Tumor cell migration is essential for tumor metastasis. Tworepresentative compounds,2-chloro-N-(6-chlorobenzo[d]thiazol-2-yl)-5-nitrobenzenesulfonamide(Compounds 3), andN-(3-(1H-1,2,4-triazol-3-ylthio)-4-hydroxynaphthalen-1-yl)-4-methoxybenzenesulfonamide(Compound 10), shown in Table 1, were selected to investigate theireffects on tumor metastasis in animal models. Tumor cells (4T1 breasttumor cells) were injected into the mammary fat-pad of mice. Themetastasis of these breast tumor cells from the mammary gland to thelung was monitored by the clonogenic assay. As shown in FIG. 1, bothrepresentative fascin inhibitors, Compounds 3 and 10, decreased thetumor metastasis in a mouse model.

Balb/c mice were purchased from Charles River. All animal procedureswere approved by the Animal Care and Use Committees of the Weill CornellMedical College and performed in accordance with institutional polices.For xenograft tumor metastasis studies, 5×10⁵ 4T1 cells were suspendedin 100 μL PBS and injected subcutaneously into the mammary glands of 6-8week old female Balb/c mice. Tumor incidence was monitored for 21 daysafter injection. Tumor size was measured three times a week, and thevolume was calculated using the formula length×width²×0.5. Compoundtreatment was initiated 7 days after tumor implantation; animals wereadministered daily with indicated dose for 2 weeks. On day 28, the micewere sacrificed. Numbers of metastatic 4T1 cells in lungs weredetermined by the clonogenic assay. In brief, lungs were removed fromeach mouse on day 28, finely minced and digested for 2 h at 37° C. in 5mL of enzyme cocktail containing PBS and 1 mg/mL collagenase type IV ona rocker. After incubation, samples were filtered through 70-μm nyloncell strainers and washed twice with PBS. Resulting cells weresuspended, plated with a series of dilutions in 10-cm tissue culturedishes in RPMI-1640 medium containing 60 μM thioguanine, metastasizedtumor cells formed foci after 14 days, at which time they were fixedwith methanol and stained with 0.03% methylene blue for counting. Dataare expressed as mean±S.D. and analyzed by Student's t test withsignificance defined as p<0.05.

The two compounds shown above were representative of those thatinhibited tumor metastasis and thus the compounds described herein arecontemplated to be useful for treating a condition or disorder mediatedby fascin activity and/or tumor metastasis.

Example 7: In Vivo Mouse Model for Prostate Tumor Metastasis

5- to 6-week-old male severe combined immunodeficient mice (n=20)purchased from Charles River (Wilmington, Mass.) are randomly dividedinto two groups (n=10 animals per group). In both two groups, humanprostate tumor cells PC-3Luc cells (stably transfected with luciferasegene) (2×10⁵ cells in 100 μl of Dulbecco phosphate-buffered saline [PBS]lacking Ca²⁺ and Mg²⁺) are introduced into animals by intracardiacinjection under 1.75% isoflurane/air anesthesia. Throughout the durationof the experiment, animals in group 1 receive daily testing compoundsadministered intraperitoneally (i.p.) in 0.2 mL of sterile physiologicalsaline beginning 1 week before tumor cell inoculation. In group 2(untreated control), animals receive a daily 0.2 mL i.p. injection ofthe vehicle, sterile physiological saline. Mice are serially imagedweekly for 5 weeks using an IVIS system (Xenogen Corp, Alameda, Calif.),and the results are analyzed using Living Image software (Xenogen). Forimaging, mice are injected with luciferin (40 mg/mL) i.p., and ventralimages are acquired 15 minutes after injection under 1.75%isoflurane/air anesthesia. At the end of the experiments, animals arekilled, and tissue is collected for histopathologic confirmation of bonemetastasis. It is contemplated that less bone metastasis is found ingroup 1 animals treated with a fascin inhibitory compound disclosedherein as compared with that found in group 2 animals. As such the testcompounds are useful for treating cancer, in particular, prostate tumormetastasis.

Example 8: In Vivo Mouse Model for Lung Tumor Metastasis

20 mice are divided into two groups, and 2×10⁶ A549 human lung tumorcells are injected into each mouse via the tail vein. One group istreated with a compound disclosed herein and another group is used ascontrol. After 8 weeks, the lungs are harvested, fixed, and embedded inparaffin. The number of metastatic lung nodules is counted in serialhistological sections stained with H&E. The areas of metastatic lungnodules are measured in scanned images of the H&E-stained tumor sectionsusing Paint.NET software. It is contemplated that the number and area ofmetastatic lung nodules in the treated animals are smaller than that ofthe untreated control animals. As such the test compounds are useful fortreating cancer, in particular, lung tumor metastasis.

Example 9: Treatment of Tumor Metastasis in Human

Human patients having metastatic breast cancer are administeredintravenously with a fascin inhibitory compound disclosed herein orplacebo in a randomized open-label trial. The patients are separatedinto 5 groups. Patients in each group are administered a daily dosage of0 mg (placebo), 100 mg, 200 mg, 500 mg, or 1000 mg of the compound,respectively, in 3-week cycles. The time to disease progression, overallresponse rate (ORR), duration of response, and overall survival (OS)rate are measured at the end of each cycle with known techniques. It iscontemplated that patients administered with the fascin inhibitorycompound have a longer mean or average time to disease progressionand/or duration of response, a higher mean or average overall responserate and/or overall survival rate, than patients administered withplacebo. Fewer new tumors distant from the original tumor site aredeveloped in patients administered with fascin inhibitory compound thanin patients administered with placebo. In a preferred embodiment, one ormore of the results are dose-responsive. Side effects are monitored andrecorded. As such the test compounds are useful for treating tumormetastasis in human.

REFERENCES

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EQUIVALENTS

The embodiments, illustratively described herein may suitably bepracticed in the absence of any element or elements, limitation orlimitations, not specifically disclosed herein. Thus, for example, theterms ‘comprising,’ ‘including,’ ‘containing,’ etc. shall be readexpansively and without limitation. Additionally, the terms andexpressions employed herein have been used as terms of description andnot of limitation, and there is no intention in the use of such termsand expressions of excluding any equivalents of the features shown anddescribed or portions thereof, but it is recognized that variousmodifications are possible within the scope of the claimed technology.Additionally, the phrase ‘consisting essentially of’ will be understoodto include those elements specifically recited and those additionalelements that do not materially affect the basic and novelcharacteristics of the claimed technology. The phrase ‘consisting of’excludes any element not specified.

The present disclosure is not to be limited in terms of the particularembodiments described in this application, which are intended asillustrations of various aspects. Many modifications and variations canbe made without departing from its spirit and scope, as will be apparentto those skilled in the art. Functionally equivalent compositions,apparatuses, and methods within the scope of the disclosure, in additionto those enumerated herein, will be apparent to those skilled in the artfrom the foregoing descriptions. Such modifications and variations areintended to fall within the scope of the appended claims. The presentdisclosure is to be limited only by the terms of the appended claims,along with the full scope of equivalents to which such claims areentitled. It is to be understood that this disclosure is not limited toparticular methods, reagents, compounds compositions or biologicalsystems, which can, of course, vary. It is also to be understood thatthe terminology used herein is for the purpose of describing particularembodiments only, and is not intended to be limiting.

In addition, where features or aspects of the disclosure are describedin terms of Markush groups, those skilled in the art will recognize thatthe disclosure is also thereby described in terms of any individualmember or subgroup of members of the Markush group.

As will be understood by one skilled in the art, for any and allpurposes, particularly in terms of providing a written description, allranges disclosed herein also encompass any and all possible subrangesand combinations of subranges thereof. Any listed range can be easilyrecognized as sufficiently describing and enabling the same range beingbroken down into at least equal halves, thirds, quarters, fifths,tenths, etc. As a non-limiting example, each range discussed herein canbe readily broken down into a lower third, middle third and upper third,etc. As will also be understood by one skilled in the art all languagesuch as ‘up to,’ ‘at least,’ ‘greater than,’ ‘less than,’ and the like,include the number recited and refer to ranges which can be subsequentlybroken down into subranges as discussed above. Finally, as will beunderstood by one skilled in the art, a range includes each individualmember.

While certain embodiments have been illustrated and described, it shouldbe understood that changes and modifications can be made therein inaccordance with ordinary skill in the art without departing from thetechnology in its broader aspects as defined in the following claims.

What is claimed:
 1. A method of inhibiting fascin expression or fascinactivity, comprising administering an effective amount of a fascininhibitor to a cell with fascin expression to thereby inhibit fascinexpression or fascin activity in the cell, wherein the fascin inhibitoris a compound of Formula A:

or a tautomer thereof, and/or a pharmaceutically acceptable saltthereof, wherein in Formula A: Y is N or CR; R¹ is phenyl, 5-memberedheteroaryl or 6-membered heteroaryl, wherein the phenyl, 5-memberedheteroaryl or 6-membered heteroaryl is optionally substituted with 1 to3 R⁶; R is hydrogen, halo or lower alkyl; L¹ is selected from the groupconsisting —(C(R⁸)₂)_(j)—, —(C(R⁸)₂)_(q)—C(O)—(C(R⁸)₂)_(r)—,—(C(R⁸)₂)_(q)—C(O)N(R⁸)—(C(R⁸)₂)_(r)—,—(C(R⁸)₂)_(q)—N(R⁸)C(O)—(C(R⁸)₂)_(r)—,—(C(R⁸)₂)_(q)—N(R⁸)S(O)₂—(C(R⁸)₂)_(r)—,—(CH₂)_(q)—S(O)₂N(R⁸)—(CH₂)_(r)—, —S—, —O— and —NR⁸—; j is 1, 2 or 3; qis 0 or 1; r is 0 or 1; L² is selected from the group consisting acovalent bond, —C(O)N(R⁸)—, —N(R⁸)C(O)—, —N(R⁸)S(O)₂—, and —S(O)₂N(R⁸)—;R⁵ is phenyl, 5-membered heteroaryl, 6-membered heteroaryl, 5-memberedheterocycloalkyl or 6-membered heterocycloalkyl; wherein the phenyl issubstituted with 1 to 4 R², and the 5-membered heteroaryl or 6-memberedheteroaryl is optionally substituted with 1 to 4 R², wherein each R² isindependently selected from the group consisting of lower alkyl, lowerhaloalkyl, —OH, —OR⁷, —SH, —SR⁷, —NR¹⁰R¹⁰, halo, cyano, nitro, —COH,—COR⁷, —CO₂H, —CO₂R⁷, —CONR¹⁰R¹⁰, —OCOR⁷, —OCO₂R⁷, —OCONR¹⁰R¹⁰,—NR¹⁰COR¹⁰, —NR¹⁰CO₂R¹⁰, —SOR⁷, —SO₂R⁷, —SO₂NR¹⁰R¹⁰, and —NR¹⁰SO₂R⁷;each R⁶ is independently selected from the group consisting of halo andlower alkyl optionally substituted with 1-3 halo; or two adjacent R⁶ ona phenyl ring form a 5- or 6-membered cycloalkyl or heterocycloalkylfused with the phenyl ring; R⁷ is lower alkyl; R⁸ is hydrogen; and eachR¹⁰ is independently hydrogen or lower alkyl, or two R¹⁰ together withthe atom(s) attached thereto form a 4- to 6-membered ring, with theproviso that when R⁵ is phenyl and L² is a covalent bond, then R⁵ issubstituted with 1 to 4 R².
 2. The method of claim 1, wherein thecompound of Formula A is selected from the group consisting of

or a tautomer thereof, and/or a pharmaceutically acceptable saltthereof, wherein n is 1, 2, 3 or 4, u is 1, 2 or 3, and L¹, and R⁶ areas defined in claim
 1. 3. The method of claim 1, wherein R¹ is phenylsubstituted with one, two, or three groups chosen from halo and loweralkyl.
 4. The method of claim 1, wherein R¹ is triazole.
 5. The methodof claim 1, wherein L² is —N(R⁸)S(O)₂—.
 6. The method of claim 1,wherein L¹ is —S—.
 7. The method of claim 1, wherein X¹ is OH and X² isO.
 8. The method of claim 1, wherein R² is independently selected fromthe group consisting of OH, halo, lower alkyl, and —OR⁷.
 9. The methodof claim 2, wherein the compound of Formula A has the structure:

or a tautomer thereof, and/or a pharmaceutically acceptable saltthereof.
 10. The method of claim 1, wherein, in the compound of FormulaA, Y is N.
 11. The method of claim 1, wherein, in the compound ofFormula A, R¹ is a 5-membered heteroaryl or 6-membered heteroaryloptionally substituted with 1 to 3 R⁶.
 12. The method of claim 1,wherein, in the compound of Formula A, L² is selected from the groupconsisting —C(O)N(R⁸)—, —N(R⁸)C(O)—, —N(R⁸)S(O)₂—, and —S(O)₂N(R⁸)—. 13.The method of claim 1, wherein, in the compound of Formula A, R⁵ is a5-membered heteroaryl, 6-membered heteroaryl, 5-memberedheterocycloalkyl or 6-membered heterocycloalkyl.